Thursday, April 08, 2021

Israeli Company Develops Faster, More Comprehensive Tuberculosis Test

Israeli tech company BATM Advanced Communications Ltd. announced on Monday that they have received full funding to develop a new rapid molecular diagnostics test for Tuberculosis (TB), which is expected to begin its testing and validation phases in the second half of 2021.

The testing and validation phases will be fully funded by Stop TB Partnership, an international alliance which is comprised of over governmental and non-governmental organisations in more than 100 countries, all dedicated to eliminating the disease.

While current gold standard tests take days to return results and have issues determining antibiotic resistance, BATM's new test returns results within approximately two hours, identifying the strain and its resistance to antibiotics. 

It does this using a combination of a one-step PCR test, developed by BATM subsidiary Adaltis, with a NATlab instrument, which uses a new isothermal RCA process developed by BATM's associate company, Ador Diagnostics. 

The testing and validation of the test will take place at the University of Heidelberg, with BATM anticipating it will officially begin selling them next year.

BATM CEO Dr. Zvi Marom said the company was delighted about the new testing method and the funding and research announcements, saying they believe it "will be particularly crucial in preventing the spread of drug-resistant TB strains." 

"We also expect our solution to be more affordable and accessible than those used today," he said.

Dr. Marom elaborated, saying that "The devastating health, social and economic consequences of TB have been with us for centuries and are still here today. It is vital that we develop innovative solutions and systems that will enable this disease, which is a leading cause of death but is both preventable and curable, to be eradicated." 

He said that advances in molecular diagnostics have caused science to enter "a new era in the fight against infectious diseases." 

"I believe that BATM has an important role to play in this as we focus on developing new, innovative technologies while continuing to provide critical solutions to combat COVID-19," he concluded.

TB is one of the top 10 causes of death worldwide and the leading (non-pandemic) cause from a single infectious agent The WHO estimates that around three million cases go undiagnosed each year.

Drug-resistance, which can emerge through the prescription of incorrect treatment, in TB is an increasing public health issue and a health security threat.  TB can also cause other severe illnesses, such as meningitis, and can be an early indicator for HIV infection.

However, "with rapid and accurate diagnosis," BATM says, "TB is preventable, treatable and curable in most cases."

MatMaCorp Launches MYRTA, a New Hand-Held Real-Time PCR Device for POC and OTC Molecular Diagnostics

MatMaCorp (Materials and Machines Corporation), a developer of comprehensive molecular diagnostic systems, today announced the launch of its new hand-held device capable of conducting polymerase chain reaction (PCR) amplification and real-time fluorescent detection, anytime, anywhere. The device, called MYRTA™ (MY Real-Time Analyzer) was developed to provide portable, PCR-based molecular diagnostic solutions for human and animal health.

“The COVID-19 pandemic has highlighted the substantial need for over-the-counter and point-of-care testing to detect viruses, like SARS-CoV-2, and other disease-causing agents, at any time and any location in the world,” said Dr. Abe Oommen, MatMaCorp founder and President. “Almost all of the current handheld molecular diagnostic solutions have focused on isothermal amplification methods, but real-time PCR is still the gold standard for diagnostics that require high sensitivity and specificity.”

Real-time PCR is the primary method to reliably detect active infection of SARS-CoV-2 (the coronavirus that causes COVID-19) in patient samples. Efforts at developing easy-to-use and affordable over-the-counter (OTC) and point-of-care (POC) devices have been constrained by the fact that RNA or DNA isolation from samples must be integrated into the PCR amplification and detection steps. This has resulted in PCR-based methods for OTC and POC devices being side-lined, regardless of the fact that real-time PCR is the benchmark by which other molecular tests are measured. MatMaCorp successfully overcame these issues in the development of MYRTA, which can simultaneously conduct PCR amplification and rapid, real-time fluorescence detection.

“The ability to detect nucleic acids in real-time while running PCR amplification directly from the sample is a game changer in molecular diagnostics,” said Dr. Oommen. “Our flexible, robust, and portable MYRTA device will enable the use of PCR in numerous situations, making point-of-care and over-the-counter PCR-based testing possible in any part of the globe. Remote hospitals and clinics, as well as the military, can use this device to monitor disease causing pathogens.”

MatMaCorp has demonstrated accurate detection of SARS-CoV-2 and influenza A and B viruses directly from samples. In addition to human health applications, the MYRTA system could also be used in animal health, as shown by its ability to detect porcine reproductive and respiratory syndrome virus (PRRSV), a virus that causes major respiratory disease in pigs.

MatMaCorp plans to submit the MYRTA system to the U.S. Food and Drug Administration’s (FDA) for consideration of Emergency Use Authorization (EUA) for COVID-19 in the coming months. The FDA previously granted EUA to MatMaCorp’s COVID-19 2SF RNA test for the detection of SARS-CoV-2 on the company’s Solas 8 portable detection system.

binx health Receives FDA CLIA Waiver for Chlamydia and Gonorrhea Test

binx health, a population health technology company that provides convenient healthcare solutions, announced today that the US Food and Drug Administration (FDA) has granted Clinical Laboratory Improvement Amendments (CLIA) waiver for the binx io system, a first-of-its kind molecular point-of-care testing platform capable of delivering central laboratory quality results in about 30 minutes, for the detection of chlamydia (CT) and gonorrhea (NG). The platform was previously 510(k) cleared by the FDA for testing male and female samples in moderate and high-complexity locations.

"With ever increasing sexually transmitted infection (STI) rates, point-of-care and CLIA-waived platforms like the binx io are essential additions to our STI control toolbox, which will increase accessibility and decrease the burden on traditional healthcare settings," said Barbara Van Der Pol, PhD, MPH, Professor of Medicine and Public Health at University of Alabama at Birmingham.

CLIA waiver clearance allows binx to facilitate single visit testing for CT/NG in any of the more than 220,000 locations across the Unites States holding a CLIA certificate of waiver, including convenient and accessible locations such as OBGYN and primary care offices, urgent care facilities, community health clinics, STD clinics and retail settings. Single-visit diagnosis and treatment have the potential to improve treatment compliance and reduce STI transmission across communities.

"Clinicians on the front lines of sexual health and wellness have long needed options for rapid diagnostic tools to address the epidemic growth in STIs. In a healthcare landscape where consumer convenience and rapid answers are an imperative, the binx io is the first chlamydia and gonorrhea test to be used in near patient settings combining the features essential to meet these needs in a critically important area of sexual health," said Jeffrey Luber, binx Chief Executive Officer. "The io Instrument's demonstrated clinical effectiveness, ease of operation, and patient convenience make it a much-needed tool with transformative implications for public health, especially now during the COVID-19 pandemic where STI prevention services nationwide have been dramatically reduced or cut altogether as resources have been allocated to focus on the COVID response. We are grateful to the countless scientists, physicians, and key opinion leaders who played an important role in today's watershed event."

The CDC estimates that 1 in 5 people in the U.S.1 have an STI; based on CDC and USPSTF guidelines There are an estimated 108M people in the United States, for whom regular STI testing is appropriate.2 3

Today, nearly all CT/NG tests are processed at central laboratories, with a delay between diagnoses and treatment in many cases of several days or more, which often results in infected patients not returning for treatment. Data show that treatment delays may lead to onward transmission and serious health consequences particularly in women, with 10-15% of un-treated women experiencing Pelvic Inflammatory Disease and nearly 30,000 cases of infertility annually in the US related to undetected chlamydia4. Due to these delays, and potentially detrimental follow-on health effects, published data5 also show that clinicians are more likely to presumptively treat symptomatic patients with antibiotics while the patient is in the clinic rather than wait for central laboratory test results. Inappropriate antibiotic treatment can promote antimicrobial resistance and can contribute to the onward transmission of the untreated infection.

The CLIA waived status allows for expansion by permitting the binx io, through its national commercial distribution partnership with McKesson, to be placed in the over 220,000 CLIA-waived locations nationwide.

IIT Delhi Develops Handheld Device For Early And Rapid Diagnosis Of Dengue

Dengue is a serious global health concern with a large population around the world facing the risk of getting infected. Early diagnosis of dengue is the key to prevent deterioration of a patient’s health. However, conventional diagnostic tools like nucleic acid detection using Reverse Transcriptase Polymerase Chain Reaction (RT-PCR) is a time taking proess and it also requires expensive equipments and reagents for the diagnosis of dengue.

These types of common diseases require a rapid, scalable and point-of-care diagnosis to be implemented at the community level and to reduce the workload of healthcare professionals. Understanding this, the GLancing Angle Deposition (GLAD) research group at IIT Delhi’s Physics Department has developed a handheld Surface Enhanced Raman Spectroscopy (SERS) based platform for early diagnosis of dengue virus. It also gives dengue test results within one hour (rapid diagnosis).

The handheld device has been succsessfully tested on the clinical blood samples collected from hundreds of individuals in collaboration with ICMR-National Institute of Malaria Research (NIMR), New Delhi.

Principal Investigator of the project and Professor in the Department of Physics, IIT Delhi, Prof. J.P. Singh’s research group is known for the synthesis of nanoscultptured thin films using a specialized technique called GLAD. This technique was employed to fabricate silver nanorods array based SERS active biosensors in order to detect pathogens. Only 2 microlitres of a diluted serum was dropcasted on the SERS substartes and Raman spectra were collected by flashing 785 nm laser beam through the device. All the data were fed in a statistical tool principal component analyzer (PCA) software. The intergrated device was able to clearly differentiate the three sets of blood samples; dengue positive, negative and healthy. This method provides a sensitive, rapid, and field deployable diagnosis of dengue at the early stage.

The research work was funded by IMPRINT India program of the Ministry of Education with New Age Instruments and Materials Pvt Ltd as the industry partner. The research was published in the journal Analytical Chemistry 92, 2527-2534 (2020).

The detection and distinction of human immunodeficiency virus (HIV-1) was also carried out in collaboration with ICMR-National AIDS Research Institute (NARI), Pune through the handheld SERS based platform. Binding of viruses directly on Ag nanorods without using antibodies or intermediate reagents was successfully demonstrated. The SERS platform was capable of distinguishing different tropic strains of HIV-1 suggesting tropism-based detection. The SERS based platform gives HIV-1 test results within an hour.

Speaking of the hand-held device, the project’s PI, Prof. J.P. Singh, IIT Delhi said, “This ultrasensitive and handy device has wide range of applications in the early-stage on-site detection of viral diseases and can produce the final report of investigation within an hour.” The research work was published recently in journal Colloids and Surfaces B: Biointerfaces 198, 111477 (2021)

Ultra-sensitive and Rapid Diagnostic Developed to Detect Ebola

An interdisciplinary team of scientists at Duke University has developed a highly sensitive and rapid diagnostic test for Ebola virus (EBOV) infection. In monkeys infected with Ebola, this diagnostic, called the D4-assay, proved to be 1000 times more sensitive than the currently approved rapid diagnostic test and capable of detecting the virus a full day earlier than the gold standard polymerase chain reaction (PCR) test.

This work, which appears in Science Translational Medicine on April 7, was done by biomedical engineers, molecular biologists, and immunologists at Duke University, and the University of Texas Medical Branch in Galveston and the Galveston National Laboratory.

Ebola virus gained global notoriety in early 2014 after an outbreak began spreading across the populous capital cities in Guinea, Liberia and Sierra Leone in Western Africa. By the time the pandemic was officially declared ended in 2016, more than 28,000 people had been infected and more than 11,300 people had died of the virus.

As of March 2021, there are two ongoing Ebola outbreaks in the Democratic Republic of the Congo that have resulted in more than 2,200 deaths, and Guinea declared a new outbreak of Ebola virus disease (EVD) after a cluster of cases appeared in February.

Early treatment and contact tracing of Ebola virus disease is critical for two reasons: First, patients are highly contagious once they begin showing symptoms, and early diagnosis can help contain the spread of EVD by make contact tracing and patient isolation easier. Second, the fatality rate of EVD can be as high as 90 percent if left untreated or treated late, but can be reduced to roughly 10 percent with monoclonal antibodies if patients are treated early in the infection.

The current gold standard for detecting Ebola virus is the reverse transcriptase polymerase chain reaction (RT-PCR) test, which identifies and amplifies viral RNA. Although RT-PCR has proven to be sensitive and capable of diagnosing Ebola infection much earlier than current alternatives, it has been difficult to implement in the remote settings where EVD outbreaks frequently originate.

Recent advances in RT-PCR test design have made it easier for untrained technicians to use the platform and reduced the need for expensive equipment. However, at $22.50 per test, the platform is expensive and, because it is lab-based, can be slow. In some studies, the median time required to confirm a diagnosis in the field is 6 days, making it a suboptimal tool when dealing with a highly contagious virus.

The current alternate diagnostic option is lateral flow assays, which detect antibodies or antigens that appear during an infection. These tests are cheaper, easy to use, and typically yield results in less than 30 minutes, but the trade-off is a greatly diminished sensitivity, so there is a greater possibility of the test missing an early infection.

To help address the shortcomings of both tests, the Duke team adapted the D4 assay, an ultrasensitive, point-of-care diagnostic previously developed by the Ahsutosh Chilkoti lab, to quickly and accurately detect secreted glycoprotein (sGP), a biomarker produced by the Ebola virus during the early stages of infection.

Besides Chilkoti, the Alan L. Kaganov Distinguished Professor of Biomedical Engineering, the team included Cassio Fontes, a former graduate student and now a senior research scientist in the Chilkoti lab, Michael Gunn, MD, a professor of medicine and immunology, and Barbara Lipes, an assistant professor of medicine at the Duke University School of Medicine.

"Prior studies suggested that Ebola virus produces secreted glycoprotein at high levels early in infection to act as a decoy and distract the immune system while the virus replicates and binds to the host cells," Fontes said. "We thought that if we could detect that, we could help facilitate earlier diagnosis, containment, and treatment during Ebola outbreak."

However, antibodies against sGP were not available, so Lipes, an antibody engineering expert in the Gunn lab, created a large library of antibodies that bind to sGP and screened for the antibodies that bound most strongly.

The two labs then worked together to identify the capture-detection antibody pair that provided the greatest sensitivity on the D4 platform.

The D4 assay for Ebola is made by inkjet printing these two antibodies against secreted glycoprotein onto a glass slide: detection antibodies, which are tagged with a fluorescent marker, and capture antibodies, which are primed to bind the target antigen. When a drop of blood or a throat or nose swab is placed on the slide, the detection antibodies separate from the array and bind to the target in the sample. These antibody-biomarker complexes then attach to the capture antibodies on the slide, which glow to indicate a capture.

To further improve sensitivity, the D4 assay is printed on a stealth polymer brush coating developed by the Chilkoti lab, which prevents non-target proteins from attaching to the slide's surface. This removes any 'background noise' on the chip, making it easier to detect very low levels of the target proteins, which makes the D4 assay very sensitive.

In parallel, the Chilkoti lab also developed a cheap but highly sensitive hand-held detector -- the D4Scope -- to read the results of the D4 assay.

"Jason Liu, a graduate student in my lab, worked to develop the D4Scope, which is a low-cost, battery-powered, compact, wide-field fluorescence reader that can image microarray spots with high sensitivity," Chilkoti said. "It was specifically designed using off-the-shelf components, so that parts could easily be replaced if the reader was damaged in the field."

To test their platform, the team worked with collaborators at the University of Texas Medical Branch in Galveston, where they showed that the D4 assay could detect EBOV in non-human primates a full day earlier than the RT-PCR.

"We've put the lateral flow assay to shame, sensitivity wise," said Fontes. "This is exciting because by understanding the biology of this virus, we've shown that there may be a target to look for where immunoassays like the D4 can outperform the PCR. It's really breaking new ground."

As the team moves forward, they aim to continue to refine their platform with the goal of shortening the time to results from 60 minutes to 30 minutes. Experiments are underway using molecular evolution to increase the sensitivity of the capture and detection antibodies.

They are also improving the design of the platform to make it a fully self-contained test. This would mean that users would only need to add a drop of blood to one port of a flow cell and a buffer solution to a second port to run the test under gravity flow. These changes would also allow for improved biocontainment and biosafety when handling potentially dangerous fluids.

Once these changes are made, the team hopes to coordinate clinical experiments in the field.

"I think it's a tremendous opportunity to really change the way Ebola testing works," Gunn said.

"When controlling an outbreak, it's key to identify the infected and be able to trace their contacts very quickly, and our hope is that test would allow you to do that," Gunn said. "With such a simple and rapid assay, you can also quickly screen people who are at risk of having contracted Ebola. It seems simple, but in the grand scheme of things could represent the difference between an outbreak and a pandemic."

This research was funded by the National Institutes of Health (NIH, R01AI150888; U19AI142785; UC7AI094660), the US Special Operations Command (w81XWH- 16C-0219), the National Council for the Improvement of Higher Education and the Science Without Borders project. Partial support was provided by the Department of Health and Human Services. The team also received the support of the Biosafety Level 4 (BSL-4) lab at the Galveston National Laboratory.

Competing Interests: The underlying technology of the D4 was developed by Angus Hucknall and Ashutosh Chilkoti, and was acquired by Immucor Inc (Norcross, GA) in 2014. The other authors declare no competing financial interests.

REFERENCE

"Ultrasensitive Point-Of-Care Immunoassay for Secreted Glycoprotein Detects Ebola Infection Earlier Than PCR," Cassio Fontes, Barbara Lipes, Jason Liu, Krystle Agans, Aiwei Yan, Patricia Shi, Daniela Cruz, Garrett Kelly, Kelli Luginbuhl, Daniel Joh, Stephanie Foster, Jacob Heggestad, Angus Hucknall, Maiken Mikkelson, Carl Pieper, Roarke Horstmeyer, Thomas Geisbert, Michael Gunn, Ashutosh Chilkoti. Science Translational Medicine, April 7, 2021. DOI 10.1126/scitranslmed.abd9696

Friday, March 19, 2021

Microtox® BT Evaluation of SARS-CoV-2 from Breath Test with Clinical Samples

DeepVerge, the environmental and life science AI company, announces initial data for ongoing Phase III clinical studies on the detection of SARS-CoV-2 on breath samples and identification of confirmed COVID19 positive patients.

Since Q3 2020, DeepVerge scientists have been working under laboratory conditions with the Spike Protein (“S-Protein”) of SARS-CoV-2 on the SARS-CoV-2 virus inside the Containment Level 3 (“CL3”) laboratories at the University of Aberdeen. In these studies, they have detected and identified the virus S-Protein in quantities at 40 femtogrami per millilitre (“Fg/mL”) at close to 100% sensitivity and specificity on DeepVerge’s Microtox® BTii nano-optofluidic chip.

In addition, under the clinical trial supervision of the Royal College of Surgeons, Ireland, 40 subjects, 16 of which were independently confirmed as COVID19 positive with PCRiii tests, provided breath samples that were tested on the Microtox® BT nano-optofluidic chip surface with Affimer® reagents (“Avacta Group”)iv and Optimers (“Aptamer Group”)v together the (“Binding Agents”).

The breath samples detected binding on the nano-optofluidic chip with a secondary antibody to the Spike Protein which was initially selected for the isolated spike protein work. Detection of the live virus was confirmed indicating 9 times increase in the digital spectrum signal on the Microtox® BT when compared to controls of nano-optofluidic chips with binding agent; and 19 times increase in signal with nano-optofluidic chips without binding agents. Additional digital background noise was indicated due to the non-specific binding of the antibody. Further data is required to confirm the same high sensitivity and specificity is achieved on breath test clinical trials which are underway.

Gerard Brandon CEO of DeepVerge plc commented:

“DeepVerge scientists have transformed its AI based water contamination detection system, developed over five years for e.coli, into the breath condensate Microtox® BT unit. Having successfully completed Phase I testing on the Spike Protein and Phase II studies with SARS-CoV-2 virus in the safety of CL3 laboratories, the initial results of Phase III real-world clinical studies in COVID19 patients have reached a major milestone with the demonstration that our Microtox® BT can deliver results in under 60 seconds from breath samples.

“The requirement for the UK Target Product Profile (“TPP”) Rapid Breath Test requires 150 confirmed positive samples and 250 confirmed negative samples. Additional supervised breath test clinical trials from a larger group is expected to provide sufficient data to meet the desired and acceptable criteria in the TPP to roll out the COVID19 and other pathogen breath tests later this year.”

Tracking progress of the stages of the infection

The Company notes the publicationvi by UK Medicines & Healthcare products Regulatory Agency (“MHRA”) of the “Target Product Profile Rapid Breath Tests for the direct and indirect detection of SARS-CoV-2”. Microtox® BT satisfies many of the “Desired” and “Acceptable” criteria within the document.

Subject to the limitations of the Binding Agents’ ability to capture the virus, the Microtox® BT breath test does or does not see the virus, eliminating false positives and enabling each test the potential to predict the following conditions:

• Asymptomatic and non-infectious,

• Asymptomatic and infectious,

• Symptomatic and infectious, and

• Symptomatic and non-infectious

Point-of-Care makes it possible to track and trace the progress of the stages of any infection, including COVID19, subject to the type of pathogen (bacteria, virus, fungi or parasite) or biomarker of a disease being targeted by the Binding Agents.

AI algorithms are designed to assess the risk of steric hindrance, in the case of SARS-CoV-2, by the capture of one S-Protein and one viral particle, blocking the binding of other viral particles in the immediate vicinity.

With ability to detect and identify the binding of individual S-Proteins at Fg/mL, the viral particle can be calculated to generate a bigger shift in the laser signal. Using AI, this relates back in Fg/mL of S-Proteins which indicates a viral load for each test subject.

The joint development program of work on the PBM-HALETM breath condensate device from PulmoBioMed is ongoing using multiplex bio-marker binding agents to analyse breath for 40 other diseases which include cancer, neurodegenerative, respiratory and metabolic conditions.

Winers Selected in XPRIZE Rapid Covid testing Competition

XPRIZE, the world’s leader in designing and operating incentive competitions to solve humanity’s grand challenges, is pleased to announce today the five winning teams in the $6M XPRIZE Rapid Covid Testing competition, with each winner creating high-quality, affordable COVID-19 testing to help society safely reopen and return to everyday activities. 

Chosen by an independent panel of judges, the grand prize winning solutions are radically affordable compared to what is currently available on the market; and are comparable to commercial offerings at measuring sensitivity, specificity and limit of detection, with a maximum turnaround time of 12 hours from sample to result

The winning teams are: 

Reliable LFC, LLC, Antigen Testing, Carlsbad, United States

ChromaCode, RNA Testing, Carlsbad, United States

Mirimus, RNA Testing, Brooklyn, United States

La Jolla Institute for Immunology, RNA Testing, La Jolla, United States

Alveo Technologies, RNA Testing, Alameda, United States

“We are thrilled to announce the winners of the XPRIZE Rapid COVID Testing, which awarded multiple winners with unique testing solutions to help prevent future supply chain problems,” said Anousheh Ansari, CEO of XPRIZE. “We started this journey to ensure communities across the globe have access to fast, affordable, and easy-to-use COVID-19 tests. We are grateful to have the best entrepreneurial and scientific teams on board to help bring their solutions to scale so we can properly reopen schools, businesses, and other vital institutions around the world.”

Following the December finalist announcements, the 20 teams sent their testing kits and protocols to two separate laboratories, for clinical validation. The independent judges, composed of diverse experts in the healthcare and COVID-19 space, reviewed each team’s lab results, testing concepts, and proposals before deciding on the winners. 

“While vaccines are important, we cannot rely on them alone to prevent the spread of the coronavirus and future outbreaks, especially not until they are provided around the world, en masse and at-scale,” said Jeff Huber, President & Co-Founder of OpenCovidScreen. “These technological breakthroughs in rapid covid testing are providing a safety net to ensure the spread of the disease is contained and to enable a safe return to work and school, and to protect hotspots like nursing homes. These advancements are key to helping underserved, under-resourced communities get access to affordable, accurate tests and to ultimately save more lives now and in the future.”

Additionally, four other teams were selected as winners in the Open Innovation Track, whose approaches demonstrated high potential for impactful screening solutions, but could not be categorized as polymerase chain reaction (PCR), Isothermal Amplification, Next Generation Sequencing, or Antigen Detection and could not be tested through the competition rounds. 

The four winning teams in the Open Innovation Track are: 

Steradian Technologies, Inc., Houston, United States

U-smell-it, Guilford, United States

Ram Global, Zweibrücken, Germany

TeraGroup, Herzliya, Israel

The XPRIZE Rapid Covid Testing judges included:

Dr. Rick Bright, Ph.D, Immunology and Molecular Pathogenesis

Shawna Butler, R.N. M.D.A., Nurse Economist

Dr. Charity Dean, CEO and Co-Founder, The Public Health Company

Dr. Paul Drain, Associate Professor, Departments of Global Health, Medicine and Epidemiology at the University of Washington

Dr. Anita Goel, Physicist and Physician, Chairman and CEO, Nanobiosym

Dr. Michael Mina, Physician-Scientist and Assistant Professor, Epidemiology and Immunology and Infectious Diseases at the Harvard School of Public Health

Dr. Anne Wyllie, Associate Research Scientist, Yale School of Public Health

“The competition was open to all modalities of molecular testing, and the teams submitted an impressive range of ideas. The winners created innovative technologies in rapid PCR, novel antigens, and point-of-care LAMP as well as pioneering some of the first-ever olfaction and breathalyzer tests,” said Chris Mason, Leader of the Science Team and a Professor at Weill Cornell Medicine.

Launched this past July amid the COVID-19 pandemic, this prize comes out of the XPRIZE Pandemic Alliance to bring researchers, innovators, institutions, corporations, and governments together to share ideas and resources in the fight against the current and future pandemics. Since launch, 85 organizations have joined the Alliance, where they have been able to share ideas and research through its digital collaboration platform Exchange, as well as through the XPRIZE Data Collaborative, a unique platform for innovators to collaborate, share and learn from data in a broad spectrum of fields in their search for solutions. 

To amplify impact, a $50 million COVID Apollo Project led by experienced life sciences investors and company builders – including RA Capital, Bain Capital, Perceptive Advisors, Redmile Group, and Samsara Biocapital – will work with OpenCovidScreen, the XPRIZE community, and beyond to accelerate the best ideas, technologies, and innovations to market and scale them.

The Anthem Foundation and Anthem, Inc., serve as the Founding Anchor Partners of XPRIZE Rapid Covid Testing. Seven major national and regional health plans are collaborating as founding partners: Blue Shield of California, Cambia Health Solutions, Inc, Health Care Service Corporation, GuideWell Mutual Holding Corporation, Horizon Healthcare Services, Inc. (NJ), BlueCross BlueShield of South Carolina. Supporting partners include leading healthcare, laboratory and technology companies: Google, Amazon, Ilumina, Ancestry, Testing for America, Thermo Fisher Scientific, Exact Sciences, Centerview Partners, Twist Bioscience, Opentrons, HudsonAlpha Institute for Biotechnology, Weill Cornell Medicine , Biotia, Inc and Medical College of Wisconsin.

Teams will work the remainder of the year to accelerate the adoption of their solutions on a massive scale. XPRIZE will oversee the development of a multimedia playbook documenting the testing protocols,plans implemented and lessons learned at deployment sites where this incredible testing technology will be rolled out. XPRIZE is currently inviting communities like schools, offices, factories, nursing homes, homeless shelters, and other communities to apply to be part of this innovative roll-out. 

Scientists Evaluate Handheld DNA Sequencers for Microbial Monitoring of Food

Researchers have evaluated a handheld DNA sequencing device for use in environmental monitoring at food factories.

The study, by researchers from the Teagasc food research program and APC Microbiome Ireland’s Science Foundation Ireland Research Centre, tested portable DNA sequencers as a routine microbial monitoring tool in food production facilities. It was funded by the Department of Agriculture, Food and the Marine (DAFM).

Small, portable, DNA sequencers provide the first steps toward real-time industry paced microbial classification and analysis, said researchers in the journal npj Science of Food.

Microbes in food can cause spoilage and disease, so routine checks in production sites are necessary. Accurate identification of microorganisms in the food chain allows sources of contamination to be identified and control measures to be put in place. However, current techniques, while tried and tested, have some limitations, researchers reported.

Advancing microbiology

“Microbiology testing in the food chain has, and continues to, rely on older, classical microbiology testing such as the use of agar and petri dishes. This is a time-consuming approach and only microorganisms that are being specifically tested for are identified,” said professor Paul Cotter, the study’s senior author.

Instead of culturing bacterial samples in petri dishes, DNA sequencing can rapidly analyze bacterial DNA and identify the species in a sample. However, conventional sequencing involves expensive lab-based equipment and only highly trained technicians can do the procedure and analyze results. This means it isn’t a good fit for routine microbial surveillance in busy food production plants.

Professor Cotter and colleagues, led by Aoife McHugh, compared the performance of Oxford Nanopore Technologies and Illumina sequencing to culture-based methods for environmental monitoring of a dairy plant.

Oxford Nanopore’s MinION handheld device was similar to the larger lab-based sequencing system in terms of the number of bacterial species it could identify in samples, suggesting it has potential as a routine monitoring device in food production. However, the small device requires a minimum amount of DNA before it can function correctly.

A step toward non-experts using DNA sequencing tools

Eight locations in the facility were swabbed on three different days in October, November, and December 2018, after cleaning in place but before the next round of dairy processing.

In the cleaned dairy facility there weren’t enough bacteria in many of the samples, so researchers had to perform an extra step to amplify the bacterial DNA before there was enough to analyze. They said further development of the technology may help to overcome this issue.

Researchers previously determined the ability of MinION-based rapid sequencing to correctly classify a four-strain, mock community of related spore-forming microorganisms of relevance to the dairy processing chain including Bacillus cereus, which can cause infections in humans.

Cotter said the use of DNA sequencing technologies to enhance food quality and safety can have an impact on everyday life.

“This study represents a key step toward a day when non-experts can use DNA sequencing tools to carry out microbiology testing in the food chain.”

Reference:

McHugh, A.J., Yap, M., Crispie, F. et al. Microbiome-based environmental monitoring of a dairy processing facility highlights the challenges associated with low microbial-load samples. npj Sci Food 5, 4 (2021). https://doi.org/10.1038/s41538-021-00087-2

Source: Food Safety News

Tuesday, March 09, 2021

Rutgers Develops Rapid Test to Detect New Emerging Coronavirus Variants

Rutgers researchers have designed a new rapid test that can detect all three of the rapidly spreading variants of the coronavirus in a little over one hour – much shorter than the three to five days required by current tests, which can also be more technically difficult and expensive to perform.

Details and information on easily creating and running the rapid test – which is not being patented by Rutgers because researchers believe it should be widely available to the public -- are published on the pre-print online server, MedRxiv, and is available at no charge.

The Rutgers researchers designed and clinically validated the test, which is the first to use “sloppy molecular beacon probes,” which are highly sensitive and specific DNA sequences used to detect frequent mutations in organisms.

“This rapid test was developed and tested over a few weeks in a crash program to respond to a serious public health need,” said David Alland, director of the Rutgers New Jersey Medical School Public Health Research Institute and professor and chief of infectious disease at Rutgers NJMS. “Despite our hurry to get the test completed, it performed extremely well with clinical samples in our initial studies. We are very pleased with these results and we hope that this test will help in the control of the rapidly evolving COVID-19 pandemic.”

The new more contagious variants, isolated in the United Kingdom, South Africa and Brazil, appear to be more easily transmitted, cause more severe disease and may be more resistant to some of the approved COVID-19 vaccines.

The new rapid test is easy to set up and can be adapted for labs that use varying types of equipment and methods. The Rutgers researchers said users are free to use the test as described or modify it as needed, although they strongly suggest that additional validation be done for any test modifications.

Researchers are also working to expand their test to more precisely differentiate among the three main viral variants and they expect to release a new and larger test menu along with supporting evidence in the next several weeks. Additional test modifications will be released in the future as additional variants emerge.

David Alland, Padmapriya Banada, Soumitesh Chakravorty, Raquel Green and Sukalyani Banik were co-researchers at Rutgers who helped develop the test.

UGA Researchers Develop New Rapid Test for Fusarium Watermelon Disease

Fusarium wilt, caused by a soilborne fungus, is one of the most damaging diseases of watermelons worldwide. Since it was discovered in 1894, it’s been a battle for producers to manage through crop rotation and chemical fungicides.

The fungus is persistent in both seedless and seeded varieties, and its spores can last for many years in the soil. While there are some watermelon varieties with known resistance, they are not resistant to all strains of the pathogen.

University of Georgia researchers have now developed a faster way to detect the disease’s presence and determine the races, or groups of strains, of the pathogen Fusarium oxysporum f. sp. niveum (FON) that cause the disease.

“At the microscopic level, you can diagnose Fusarium, but you can’t differentiate the races,” Emran Ali in the Department of Plant Pathology at the College of Agricultural and Environmental Sciences said. “Traditional bioassay methods have been used for this, but it takes weeks to grow watermelon plants and evaluate the disease, and watermelon cultivars used for the bioassay can be difficult to source. This method is not only inefficient, it is also sometimes inaccurate.”

The new molecular detection method, published in the International Journal of Molecular Sciences, allows differentiation of the different races of the pathogen.

Ali, who is head of the Plant Molecular Diagnostic Laboratory at the UGA Tifton campus, worked with Pingsheng Ji, a professor of plant pathology, and Owen Hudson, a graduate student in the department, as well as faculty at the University of Florida and Clemson University to identify more than 160 Fusarium samples collected from Georgia, Florida and South Carolina. Most of the results matched those done with the traditional bioassay method for the same samples.

The new process should improve the speed and accuracy of current diagnostic ability for FON races and facilitate research in related areas — especially breeders who are developing new varieties — to develop options such as host resistance against the disease.

“The whole process takes about three hours to diagnose races; traditional bioassay takes more than a month, at least,” Ali said.

Determining the races can help farmers better mitigate the issue and make the right management decisions. So far, four races of FON have been identified. Some commercial watermelon varieties are resistant to races zero and one, but not races two and three.

“Resistant watermelon varieties are effective against some races but not others,” Ali said. “If you quickly diagnose, growers can have more time in advance to know what’s going on in their fields. It’s good to know what’s going on. Watermelon varieties resistant to races zero and one are available, so you may grow resistant varieties to control disease caused by these races; other races are more destructive and more difficult to control.”

Georgia is continually a national leader in watermelon production, ranking second or third each year. The crop’s farm gate value was $180 million in 2019 and, with already thin profit margins, finding more efficient ways to battle diseases like Fusarium wilt could make a big difference for farmers.

Ali and his team at the Plant Molecular Diagnostic Laboratory also have developed faster tools for tracking other pathogens, including cucurbit leaf crumple virus in vegetables, dollar spot in turfgrass, citrus greening disease, root-knot nematodes in pecans, and irrigation water molds that can cause root rot. They say they hope these new detection methods will not only save time but also reduce pathogen prevalence in the field.

Coronavirus-like Particles Could Ensure Reliability of Simpler, Faster COVID-19 Tests

Rapid COVID-19 tests are on the rise to deliver results faster to more people, and scientists need an easy, foolproof way to know that these tests work correctly and the results can be trusted. Nanoparticles that pass detection as the novel coronavirus could be just the ticket.

Such coronavirus-like nanoparticles, developed by nanoengineers at the University of California San Diego, would serve as something called a positive control for COVID-19 tests. Positive controls are samples that always test positive. They are run and analyzed right alongside patient samples to verify that COVID-19 tests are working consistently and as intended.

The positive controls developed at UC San Diego offer several advantages over the ones currently used in COVID-19 testing: they do not need to be kept cold; they are easy to manufacture; they can be included in the entire testing process from start to finish, just like a patient sample; and because they are not actual virus samples from COVID-19 patients, they do not pose a risk of infection to the people running the tests.

Researchers led by Nicole Steinmetz, a professor of nanoengineering at UC San Diego, published their work in the journal Biomacromolecules.

Journal Reference:

Soo Khim Chan, Pinyi Du, Caroline Ignacio, Sanjay Mehta, Isabel G. Newton, Nicole F. Steinmetz. Virus-Like Particles as Positive Controls for COVID-19 RT-LAMP Diagnostic Assays. Biomacromolecules, 2021.


This work builds on an earlier version of the positive controls that Steinmetz's lab developed for the RT-PCR test, which is the gold standard for COVID-19 testing. The positive controls in the new study can be used not only for the RT-PCR test, but also for a cheaper, simpler and faster test called the RT-LAMP test, which can be done on the spot and provide results in about an hour.

Having a hardy tool to ensure these tests are running accurately -- especially for low-tech diagnostic assays like the RT-LAMP -- is critical, Steinmetz said. It could help enable rapid, mass testing of COVID-19 in low-resource, underserved areas and other places that do not have access to sophisticated testing equipment, specialized reagents and trained professionals.

Upgraded positive controls

The new positive controls are essentially tiny virus shells -- made of either plant virus or bacteriophage -- that house segments of coronavirus RNA inside. The RNA segments include binding sites for both of the primers used in the PCR and LAMP tests.

"This design creates an all-in-one control that can be used for either one of these assays, making it very versatile," said first author Soo Khim Chan, who is a postdoctoral researcher in Steinmetz's lab.

The team developed two types of positive controls. One was made from plant virus nanoparticles. To make them, the researchers infected cowpea plants in the lab with cowpea chlorotic mottle virus and then extracted the viruses from the plants. Afterwards, the researchers removed the virus' RNA and replaced it with a custom-made RNA template containing specific yet non-infectious sequences from the SARS-CoV-2 virus. The resulting nanoparticles consist of coronavirus RNA sequences packaged inside plant virus shells.

The other positive control was made from bacteriophage nanoparticles. It involved a similar recipe. The researchers infected E. coli bacteria with custom-made plasmids -- rings of DNA -- that contain specific fragments of sequences (which are also non-infectious) from the SARS-CoV-2 virus, as well as genes coding for surface proteins of a bacteriophage called Qbeta. This process caused the bacteria to produce nanoparticles that consist of coronavirus RNA sequences packaged inside bacteriophage shells.

The plant virus and bacteriophage shells are key to making these positive controls so sturdy. They protect the coronavirus RNA segments from breaking down at warmer temperatures -- tests showed that they can be stored for a week at temperatures up to 40 C (104 F). The shells also protect the RNA during the first step of the PCR and LAMP tests, which involves breaking down cells in the sample -- via enzymes or heat -- to release their genetic material for testing.

These protections are not present in the positive controls currently used in COVID-19 testing (naked synthetic RNAs, plasmids or RNA samples from infected patients). That's why existing controls either require refrigeration (which makes them inconvenient to handle, costly to ship and store) or have to be added at a later stage of the test (which means that means scientists will not know if something went wrong in the first steps).

As a next step, the researchers are looking to partner up with industry to implement this technology. The positive controls can be adapted to any established RT-PCR or RT-LAMP assay, and using them would help negate false readouts, Steinmetz's team said. Plus, these positive controls can be easily produced in large quantities by molecular farming in plants or microbial culture fermentation, which is good news for translating them to large-scale manufacturing.

"With mutants and variants emerging, continued testing will be imperative to keep the population safe," Steinmetz said. "The new technology could find utility in particular for at-home tests, which may have a higher rate of false readouts due to the less controlled experimental conditions."

This work was funded in part by the National Science Foundation (RAPID CBET-2032196 and RAPID CMMI-2027668) and the University of California (UCOP-R00RG2471 and a Galvanizing Engineering in Medicine Award).

Liquid Crystal Sensor Offers Breakthrough for Rapid COVID-19 Diagnostic Test

The global outbreak of COVID-19 has prompted scientists to focus their research on rapid and robust diagnostic tests. Reverse-transcription polymerase chain reaction (RT-PCR) assays – the ‘gold-standard’ for molecular clinical diagnostics – became available quickly, however this provides relatively long characterisation times and the need for specialised equipment.

Researchers at OSU have subsequently designed a liquid crystal (LC) based diagnostic kit and a smartphone-based application to enable automatic detection of SARS-CoV-2 ssRNA, which could be used for reliable self-test of SARS-CoV-2 at home without the need for complex equipment or procedures.

Prof Xiaoguang (William) Wang, PhD led the research team at OSU, and commented on his decision to consider LCs as a prospective diagnostic technology to meet the need: “It is well known that thermotropic LCs are ultrasensitive to small chemical modulations, and we felt that we could use this property to develop a rapid LC-based detection method for SARS CoV-2 virus that could be contained in a portable and economical sensor.”

A series of experiments were required to prove this concept and develop a prototype sensor, which included three key steps. 

1. The characterisation of a substrate layer that had been ‘functionalised’ with a liquid crystal mixture.

2. Validation of the reliable binding of an ssDNA ‘probe’ to this layer.

3. The detection and quantitation of ssRNAcov extracted from the virus as it bound to the probe.

Important for this last step was holding the temperature of the system at precisely 48.7°C, the melting temperature of ssRNA. The Linkam PE120 stage was integral to this phase and gave the team at OSU the reproducible control of the temperature of the sensor they needed during the process.

The PE120 Peltier system is straightforward to integrate with microscopes and other equipment, and ensures precise control of the temperature of microscope slides to +/-0.1°C at temperatures between  25°C to 120°C. 

Prof Wang’s research has recently been published, and it has shown that LC-based sensors offer a highly sensitive, reproducible and robust method of detection of target ssRNAcov – with detection limits in the nanomolar to femtomolar range.

Commenting on the work, Robert Gurney PhD, Marketing and Applications Specialist at Linkam Scientific, said: “What Prof Wang and his colleagues have achieved is extremely exciting. It’s great that one of the Linkam temperature control stages has played a key part in the team’s proof-of-concept studies, and I’m looking forward to seeing the results of the next stage – trials of the sensor with patient samples.”

Mobidiag Receives CE-IVD Marking for Amplidiag® RESP-4 for Simultaneous Detection of COVID-19, Flu A, Flu B and RSV

Mobidiag Ltd. today announces that it has received CE-IVD marking for its Amplidiag® RESP-4 molecular diagnostic test for the rapid and simultaneous detection of some of the most prevalent respiratory viruses: SARS-CoV-2 (responsible for COVID-19 infection), Influenza A, Influenza B and RSV.

These viruses present similar symptoms, making it difficult for physicians to determine accurately which virus a patient has. The Amplidiag® RESP-4 assay can rapidly and simultaneously determine an accurate diagnosis from one patient sample using nasopharyngeal swabs, helping to guide clinicians to the relevant treatment decision for each patient. The test runs on the Mobidiag Amplidiag® Easy platform, which provides clinicians with an optimized sample screening process, with automated DNA/RNA extraction and PCR plate setup. Based on well-established high-throughput PCR technology, the platform can process 46 samples in approximately three hours.

This new test further broadens Mobidiag’s already available portfolio of diagnostics tests.

Tuomas Tenkanen, CEO of Mobidiag said, “We are very pleased to announce the CE-IVD marking for the Amplidiag® RESP-4 assay, which is highly relevant to both the traditional flu season and COVID-19. Symptoms caused by these viruses [SARS-CoV-2, Influenza A, Influenza B and RSV] can be very similar, and it is imperative that healthcare professionals are able to rapidly distinguish between them, in order to start the appropriate treatment for the benefit of patients.

In response to the growing need for fast and reliable diagnostics, at Mobidiag we are putting all our efforts into maintaining a high degree of autonomy in the production of our Amplidiag® tests by securing critical components to provide continuous support for our customers, as we all work together to control the spread of COVID-19.”

Mobidiag is currently developing Novodiag® RESP-4, a molecular diagnostic test using its Novodiag® platform for the rapid and on-demand detection of SARS-CoV-2, Influenza A, Influenza B and RSV. This test will complement Amplidiag® RESP-4 and enable clinicians to detect respiratory viruses to support early decisions in managing efficiently epidemiological and infection control measures, isolate patients in a timely manner, and ultimately improve patient care and outcomes.

Please note that Mobidiag tests are not home testing kits. They are only available for healthcare professionals, and not for patients directly. Please follow the recommended processes and guidance for your location if you have any symptoms of respiratory infections.

About Mobidiag

Mobidiag is a fast-growing molecular diagnostics company whose rapid, cost-effective, widely applicable and robust technology makes the power of molecular diagnostics available to address the spread of infectious diseases and antimicrobial resistance (AMR) by rapid detection of pathogens and their potential resistance to antibiotics. Through its Amplidiag® and Novodiag® solutions, Mobidiag offers a comprehensive range of molecular diagnostic solutions for the detection of infectious diseases to laboratories of all sizes.

Tuesday, February 23, 2021

U of Minnesota Researchers Develop Two New RZapid COVID-19 Diagnostic Tests

University of Minnesota Medical School researchers have developed two new rapid diagnostic tests for COVID-19 - one to detect COVID-19 variants and one to help differentiate with other illnesses that have COVID-19-like symptoms. The findings were recently published in the journal Bioengineering.

Although many people are hopeful about COVID-19 vaccines, widespread vaccine distribution isn't predicted to be available until several months from now. Until that happens, the ability to diagnose COVID-19 quickly and accurately is crucial to help minimize loss of life and continued spread of the virus.

The technology for both tests uses the cutting-edge CRISPR/Cas9 system. Using commercial reagents, they describe a Cas-9-based methodology for nucleic acid detection using lateral flow assays and fluorescence signal generation.

The first test is a rapid diagnostic test that can differentiate between COVID-19 variants. This test can be performed without specialized expertise or equipment. It uses technology similar to at-home pregnancy testing and produces results in about an hour.

The second, more sensitive test allows researchers to analyze the same sample simultaneously for COVID-19 (SARS-CoV-2), Influenza A and B and respiratory syncytial virus by measuring fluorescence. These viruses manifest with similar symptoms, so being able to detect and differentiate them adds a new diagnostic tool to slow the spread of COVID-19. This test also takes about an hour and could be easily scaled so many more tests can be performed. The necessary equipment is present in most diagnostics laboratories and many research laboratories.

"The approval of the SARS-CoV-2 vaccine is highly promising, but the time between first doses and population immunity may be months," said Mark J. Osborn, PhD, assistant professor of Pediatrics at the University of Minnesota Medical School and first author of this paper. "This testing platform can help bridge the gap between immunization and immunity."

In collaboration with the U of M's Institute of Engineering in Medicine and Jakub Tolar, MD, PhD, dean of the U of M Medical School, Osborn and his team are now seeking to enhance sensitivity and real-world application of this test in support of rapidly detecting and identifying COVID-19 variants. In order to provide access to their new testing technology for healthcare providers and the public, the researchers are currently exploring ways to scale up and license their new diagnostics.

DARPA Backs Rice University Sensor to Detect COVID-19 Virus in Air

Researchers at Rice University have received funding for up to $1 million to develop a real-time sensor system able to detect minute amounts of the airborne virus that causes COVID-19 infection.

The researchers at Rice's Brown School of Engineering and Wiess School of Natural Sciences -- chemical and biomolecular engineer Rafael Verduzco, civil and environmental engineer Pedro Alvarez and structural virologist Yizhi Jane Tao -- will team with William Lawrence, a microbiologist at the University of Texas Medical Branch (UTMB) at Galveston to develop a thin film electronic device that senses as few as eight SARS-CoV-2 viruses in 10 minutes of sampling air flowing at 8 liters per minute.

The project titled Real-Time Amperometric Platform Using Molecular Imprinting for Selective Detection of SARS-CoV-2 (RAPID) has been funded by the Defense Advanced Research Projects Agency (DARPA). The second half of funding is contingent upon a successful demonstration of the technology.

"We had started working last summer on the idea of trying to detect SARS-Cov-2," said Verduzco, a professor of chemical and biomolecular engineering and of materials science and nanoengineering and principal investigator on the project.

"Pedro initiated the idea because he had some films that incorporate molecularly imprinted polymers that he thought could very selectively respond to anything.

"He thought we could modify it to emit an electronic signal when a virus binds to the film," Verduzco said. "Jane got involved because we want a biologist to help build these recognition layers. We saw the opportunity to pursue this with DARPA, because they have a very challenging but specific metric for sensing a very low concentration of SARS in air within 10 minutes."

Alvarez and Tao previously introduced a filter that could "trap and zap" SARS-CoV-2 in wastewater at treatment plants, a technology that was itself adapted from their method to kill bacterial "superbugs" and degrade their antibiotic resistance genes.

"Molecular imprinting cavities where specific molecules or particles fit snugly can enhance the capacity of surfaces to selectively adsorb and concentrate viruses, which in turn facilitates their disinfection, in the trap-and-zap project, or detection, in this RAPID project," Alvarez said.

"Thus, we were able to leverage previous work on molecular imprinting," he said. "Jane suggested a significant improvement related to anchoring specific biorecognition factors to further enhance the selectivity of the surface to attach and concentrate SARS-CoV-2."

The researchers' proposal describes a bioaerosol sampler that would concentrate airborne SARS-CoV-2 into a liquid electrolyte medium, bind it onto virus-imprinted polymers functionalized with SARS-CoV-2 attachment factors that enhance selectivity and use organic electrochemical transistors to rapidly transduce SARS-CoV-2 binding events into electronic signals.

The proposed device would be sized for analysis of a 50-cubic-meter office, a 300-cubic-meter classroom or central building monitoring. They expect the filtration system to be not only rapidly adaptable for other pathogens but also able to nondestructively capture viruses in a way that retains them for further analysis.

Lawrence's lab works with the UTMB Galveston National Laboratory, which is part of the National Institute of Allergy and Infectious Diseases biodefense network. He is also director of the Aerobiology Services Division at the lab and has expertise with aerosolization and testing of SARS-CoV-2.

Verduzco said the Rice team will spend the first nine months fabricating the device and testing it on inactivated SARS-CoV-2. "If we are successful, the next nine months will focus on testing with live SARS-CoV-2 at UTMB, and also optimizing the device to meet project metrics," he said.

Bio-Aerosol Collection and Identification System Capable of Detecting SARS-CoV-2 in Air

Smiths Detection, a global leader in threat detection and security screening technologies, today reports that its BioFlash® Biological Identifier is capable of detecting SARS-CoV-2 in the air following tests conducted by the United States Army Medical Research Institute of Infectious Diseases (USAMRIID).

The tests were performed using live SARS-CoV-2 virus in a Biosafety Level 3 containment area at Fort Detrick, Maryland. The SARS-CoV-2 CANARY biosensor used in the BioFlash device demonstrated that it can quickly detect and identify the presence of low levels of aerosolized SARS-CoV-2.

The BioFlash® Biological Identifier is powered by CANARY® technology (a cell-based biosensor) and is combined with proprietary aerosol-collection techniques to provide rapid, sensitive and specific identification of biological-threat agents including viruses, toxins and bacteria.

“We are working incredibly hard to provide a tool that will support the ongoing fight against the coronavirus,” said Roland Carter, President, Smiths Detection. “BioFlash is an effective and trusted environmental monitoring tool. These test results provide valuable data in understanding the spread of COVID-19 and help protect people in indoor environments such as hospitals, schools and commercial buildings.”

USAMRIID confirmed that Smiths Detection’s BioFlash can detect down to an estimated 6,000 airborne infectious particles of the SARS-CoV-2 virus within a controlled environment. This compares to as many as one million particles emitted in a single sneeze by a person infected with SARS-CoV-2. The test results also indicate no cross-reactivity with influenza and Middle East Respiratory Syndrome (MERS), an important consideration for environmental monitoring of the SARS-CoV-2 virus.

Further testing and research is underway at a number of US universities to collect more data on how the detection technology can help prevent outbreaks and guide both public and private organizations in COVID-19 mitigation strategies.

Malta Researchers to Develop a Non-Contact Hyperspectral Imaging Method to Detect Microbial Contamination in Food

A project financed by the Malta Council for Science and Technology has developed a non-contact and non-destructive approach for the early detection of microbial contaminants that are responsible for food spoilage, with a focus on slow-growing fungi in dairy products.

The MCST awarded €195,000 to the project, which involved a collaboration between the Centre for Biomedical Cybernetics, the Department of Food Sciences and Nutrition at the University of Malta, and Farm Fresh Ltd.

Every year, the European dairy industry processes approximately 152 million tons of raw milk, for consumption or for the production of food, feed and pharmaceutical products. The raw milk delivered by the EU-25’s 1.6 million dairy farmers, processed by the dairy industry, plays a vital role in rural areas, and the dairy industry represents approximately 15 per cent of the turnover of the food and drinks industry in Europe, employing about 13 per cent of the total workforce.

Typical tests currently in use for the analysis of milk products rely on lengthy procedures that can last from 24 to 36 hours for bacterial analysis, and seven to eight days for fungal analysis. Alternative methods such as rapid genomic subtyping may be faster but are very costly for SMEs not running their own research and development department, while the efficacy of methods such as infrared spectroscopy can be limited if the presence of water is above specific thresholds.

Owen Falzon, senior lecturer at the University’s Centre for Biomedical Cybernetics, said: “The FIHI project consortium investigated the use of a hyperspectral imaging to assess the characteristics of food products at different spectral bands. These images can be considered as a fingerprint that characterises the composition of the object being analysed.

“Through the automated processing and analysis of the hyperspectral data, this system can help identify contaminated products while reducing time and effort for food sample inspection.”

In light of recent food-borne illness outbreaks, the early detection of contaminated products in the processing chain can allow for immediate action to prevent contaminated batches from moving further down the production and distribution line and reaching the end customer, leading to a significant social as well as economic impact especially in regions at greater risk.

The Maltese ġbejna (cheeselet) is made from sheep or goat milk curds and aged for several months to develop its distinctive taste. During the ageing process, the cheese can become spoiled by fungi and unsafe for human consumption.

This is a significant public health risk and a financial liability for producers. Conventional microbiology techniques may involve lengthy analysis procedures for the detection of these slow-growing unpigmented fungi, allowing occasional distribution of contaminated products.

To test this hypothesis, a model cheeselet was produced with the involvement of Farm Fresh Ltd to conduct compatibility and stability studies, through measurements of colony forming units, water activity, moisture levels, pH, protein and sugar content. The ġbejna model was then challenged with fungal strains isolated from commercial ġbejna and imaged using a hyperspectral camera.

Graphene Biosensor Allows Quick Detection of Multiple Sepsis Biomarkers in Blood

A specialized biosensor allows quick and accurate detection of multiple sepsis biomarkers – procalcitonin, C-reactive protein and pathogen-associated molecular patterns — from a blood sample.

The sensor was developed as a collaboration between Harvard’s Wyss Institute for Biologically Inspired Engineering and the University of Bath in the UK. It combines graphene nanoparticles with a bovine serum albumin composite to allow the detection of multiple biomarkers in one test.

Sepsis is an inflammatory immune response that is triggered in response to an infection. It can be life threatening and patients often deteriorate fast. This means that every minute counts and a quick diagnosis is important to make sure patients are treated correctly and have the best chance of a fast recovery.

Various attempts have been made to produce fast, point of care electrochemical tests for different biomarkers, but so far these all test one biomarker at a time. Some, such as Abbott’s i‐STAT system, allow the use of different cartridges for different biomarkers, but they cannot test concurrently for more than one biomarker.

A confirmed sepsis diagnosis requires testing of multiple different biomarkers, but running several separate tests takes time. It has therefore long been a goal to develop an electrochemical sensor that can run multiple tests at the same time.

Wyss Founding Director Donald Ingber, M.D., Ph.D., and colleagues achieved this by combining graphene nanoparticles, which helps prevent biological material sticking to the sensor while maintaining electroconductivity, with a thin composite coating of bovine serum albumin. When desired biomarkers stick to the coating, an electrical signal is generated and a measurement recorded.

This is an advance on previous technology developed by the same group that used gold rather than graphene. “We replaced the coating’s gold nanowires with graphene oxide nanoflakes that also have anti-fouling and electrochemical properties, but they are much less expensive and allow even more sensitive measurements. In fact, the costs of fabricating the nanocomposite were reduced to a fraction of its original cost,” said Wyss Senior Staff Scientist Pawan Jolly, Ph.D., who also worked on the technology.

The team first added antibodies to the sensor that could detect procalcitonin, which is produced by many cells in response to bacterial infections and checked its accuracy compared with a standard ELISA assay.

They then created a multiplex sensor by adding elements that could detect C-reactive protein, a marker of inflammation often raised in sepsis, and pathogen-associated molecular patterns. The latter element uses a genetically modified protein called FcMBL, developed at Wyss, that has the ability to bind over 100 different pathogenic microbes and their associated biomarkers, which are often released into the blood during sepsis.

As reported in the journal Advanced Functional Materials, the researchers found that the multiplex sensor detected biomarkers in a clinically significant range and did not show any cross-reactivity, which has caused problems with accuracy in the past.

“Assembling three dedicated electrochemical sensor elements for biomarkers that can be present in blood at vastly different concentrations on a single chip posed a significant challenge. However, the three elements in the final sensor exhibited specific responses within the clinically significant range without interfering with each other, and they did so with a turnaround time of 51 minutes, which meets the clinical need of sepsis diagnosis within the first hour,” said Uroš Zupančič, a Ph.D. student who was previously a visiting scholar in Ingber’s group from the University of Bath.

To create a proof of concept, the team integrated the procalcitonin sensor with a microfluidic system that allows inclusion of more biomarker binding sites and adds automation. This allowed detection within 7 minutes. Although only one biomarker was tested in this way, the researchers think it should be fairly straightforward to add additional sensors in close proximity on the same chip.

“When coupled with microfluidics, these novel multiplexed sensors could detect a panel of biomarkers in whole blood in a matter of minutes, opening the way to low‐cost and easy‐to‐use rapid sepsis diagnostics,” conclude the authors.

A specialized biosensor allows quick and accurate detection of multiple sepsis biomarkers – procalcitonin, C-reactive protein and pathogen-associated molecular patterns — from a blood sample.

The sensor was developed as a collaboration between Harvard’s Wyss Institute for Biologically Inspired Engineering and the University of Bath in the UK. It combines graphene nanoparticles with a bovine serum albumin composite to allow the detection of multiple biomarkers in one test.

Sepsis is an inflammatory immune response that is triggered in response to an infection. It can be life threatening and patients often deteriorate fast. This means that every minute counts and a quick diagnosis is important to make sure patients are treated correctly and have the best chance of a fast recovery.

Various attempts have been made to produce fast, point of care electrochemical tests for different biomarkers, but so far these all test one biomarker at a time. Some, such as Abbott’s i‐STAT system, allow the use of different cartridges for different biomarkers, but they cannot test concurrently for more than one biomarker.

A confirmed sepsis diagnosis requires testing of multiple different biomarkers, but running several separate tests takes time. It has therefore long been a goal to develop an electrochemical sensor that can run multiple tests at the same time.

Wyss Founding Director Donald Ingber, M.D., Ph.D., and colleagues achieved this by combining graphene nanoparticles, which helps prevent biological material sticking to the sensor while maintaining electroconductivity, with a thin composite coating of bovine serum albumin. When desired biomarkers stick to the coating, an electrical signal is generated and a measurement recorded.

This is an advance on previous technology developed by the same group that used gold rather than graphene. “We replaced the coating’s gold nanowires with graphene oxide nanoflakes that also have anti-fouling and electrochemical properties, but they are much less expensive and allow even more sensitive measurements. In fact, the costs of fabricating the nanocomposite were reduced to a fraction of its original cost,” said Wyss Senior Staff Scientist Pawan Jolly, Ph.D., who also worked on the technology.

The team first added antibodies to the sensor that could detect procalcitonin, which is produced by many cells in response to bacterial infections and checked its accuracy compared with a standard ELISA assay.

They then created a multiplex sensor by adding elements that could detect C-reactive protein, a marker of inflammation often raised in sepsis, and pathogen-associated molecular patterns. The latter element uses a genetically modified protein called FcMBL, developed at Wyss, that has the ability to bind over 100 different pathogenic microbes and their associated biomarkers, which are often released into the blood during sepsis.

Source: Clinical OMICS

FSU Professor Receives USDA Grants to Develop Food Safety Tests Using Gold Nanoparticles

A Florida State University researcher has received two grants from the U.S. Department of Agriculture to develop tests that will uncover adulterated or contaminated foods.

“These projects will protect against contamination of the food supply that may cause wide-scale public health harm,” said Qinchun Rao, an associate professor in the Department of Nutrition, Food and Exercise Science. “They’ll also help ensure that consumers get the products they think they are paying for.”

Rao will develop an easy-to-use test that can detect whether a food contains shellfish, a potentially fatal allergen for some people. Rao also will collaborate with Xiaohu Xia, an assistant professor in the Department of Chemistry at the University of Central Florida (UCF), on a separate project to develop a test for the presence of adulterated meat.

About one-third of food recalls in the U.S. are because of misbranded or undeclared allergenic food residues. Unknown shellfish or meat might be present in food products because of accidental cross-contamination, or they might be added intentionally in place of another, more expensive ingredient. Consumers eating adulterated food might be paying extra for a tampered product, inadvertently violating religious dietary restrictions or eating something to which they’re allergic.

In the shellfish project, Rao’s team will purify four major shellfish allergens from shrimp and crabs and develop tests for those allergens.

“The long-term goal of this project is to provide a robust diagnostic tool for the enforcement of the U.S. Food and Drug Administration’s Food Safety Modernization Act in order to prevent cross-contamination of shellfish allergens in foods,” Rao said. “There’s a need for tests that are reliable but also convenient to use and inexpensive.”

As part of the project to test for adulterated meat with UCF, Rao’s lab will analyze the test strip built by Xia to determine its effectiveness. The team will improve upon existing technology that uses gold nanoparticles to detect meat proteins. Preliminary results have shown that adding a coating of platinum, palladium or iridium to the gold nanoparticles makes the test more sensitive. The researchers will continue their work to improve the testing procedure by using other metals in the nanoparticle coating.

The goal is to develop tests that are sensitive, easy-to-use and affordable, which would make them useful tools for deploying on a wide scale to find cases of adulterated food.

“The success of this project will be of great benefit to fighting food fraud by providing a simple, paper-based test strip for the rapid and sensitive detection of animal-derived adulterants in foods,” Rao said. “A newly enacted intentional adulteration rule requires mitigation strategies for processes in certain registered food facilities, and this project will help meet that requirement. In addition, these new methods being developed may open up new approaches for the food industry and the food regulatory authorities in the future.”

This work is supported by the USDA National Institute of Food and Agriculture, Agriculture and Food Research Initiative (AFRI) projects 2020-03475 and 2019-05845. A grant of about $474,000 will fund the shellfish project, and a $490,000 grant will fund the development of a test for adulterants derived from animals.

New Rapid Test Uses Magnetic Nanoparticles to Detect Coronavirus Antibodies

An international research team involving the universities of Paraná (Brazil) and Tübingen (Germany) has developed a rapid test that can reliably identify Covid-19 antibodies in the blood within minutes.

As the researchers report in the journal ACS Sensors ("Magnetic Bead-Based Immunoassay Allows Rapid, Inexpensive, and Quantitative Detection of Human SARS-CoV-2 Antibodies"), the new process is based on a simple measuring principle making it easy to carry out without expensive instruments, and is therefore suitable for use at mobile testing centers or by laboratories in less economically developed regions.

The new diagnostic method is also far faster than the enzyme linked immunosorbent assay (ELISA) procedure, which for decades has been seen as the gold standard for laboratory diagnosis of antibodies.

“Only a small sample is needed for the test: a single drop that contains two microliters of serum is sufficient,” says the lead author of the study, Professor Luciano F. Huergo from the University of Paraná: “It’s also possible to use whole blood, in other words the separation of soluble blood components that is normally necessary can be omitted.” So, the test can be used on site at care homes and testing centers. “It isn’t absolutely necessary to have a fully-equipped laboratory or use special equipment to carry it out.” In addition, the total response time is 15 times shorter than that of the classic ELISA test, as Huergo explains: “It means hundreds of samples can be tested in just a few hours.”

The new test is based on magnetic nanoparticles that are coated with viral antigens. To conduct the test blood serum or blood is applied to the test surface. After roughly two minutes the nanoparticles are washed and treated with a developer reagent. If the blood sample displays antibodies to coronavirus, a color change occurs. While the traditional ELISA test produces results after about three hours, study results show that the new method only takes twelve minutes.

Potentially useful for those who are acutely ill or recovered

Antibodies to the SARS-CoV-2 coronavirus generally form eleven to 16 days after symptoms occur. However, some patients produce detectable concentrations of antibodies as early as two to four days after the first symptoms of the disease. Therefore, immunological tests can function as additional tools to identify patients in the acute phase of Covid-19 or patients who receive a false negative from a PCR test.

“In particular for samples with low antibody titers, our test came off better than the ELISA procedure,” says Professor Karl Forchhammer from the Interfaculty Institute for Microbiology and Infection Medicine (IMIT) of the University of Tübingen. “The method worked with a sensitivity of 87 percent and a specificity of 99 percent of the tested Covid-19 samples.”

Positive and negative results can be established simply with the naked eye, and by using additional instruments, such as a microplate reader, the precision of the test can be further increased.

“Another advantage over the ELISA procedure is that the color result of our new procedure is directly proportional to the concentration of antibodies,” says Huergo. “In other words, the new method delivers data on the quantity of antibodies and not just whether any are present.”

In addition, the study shows that the new technology can also be applied to the serological diagnosis of other diseases. Professor Huergo says that the new procedure has the potential to replace the ELISA test, which has been in use since the 1970s: “We believe this technology represents a milestone in the development of immunological diagnostics.”

There are no reports in the research literature on an immunology test for Covid-19 that delivers data as quickly, as precisely and above all as cheaply.

The study authors assume that in future it will be possible to offer the new test at a comparable price to the ELISA test.

“The test only requires minimal instrumentation in all production phases and will now be evaluated with a larger number of samples and for mass production, and we believe that our fast and quantitative method for detecting SARS-CoV-2 antibodies can help to track cases of Covid-19, especially in developing countries like Brazil, who do not have the luxury of doing regular PCR-based tests, at point of care units,” says Dr. Khaled Selim, head of the German team at the University of Tübingen.

The technology is available to research, development and innovation partners via the innovation agency of the University of Paraná, which holds the statutory and patent rights.

Nanomix Submits Emergency Use Authorization Request to FDA for Rapid Antigen Panel to Detect COVID-19

Nanomix, a leader in the development of mobile, affordable, point-of-care diagnostics, today announced that the company has submitted an Emergency Use Authorization request to the U.S. Food and Drug Administration (FDA) for the eLab® COVID-19 rapid antigen test. The assay runs on the portable Nanomix eLab analyzer, which provides results in just 15 minutes and can be used in a wide range of settings, including hospitals, nursing homes, assisted living facilities, urgent care centers, and emergency medical care.

The Nanomix eLab platform and COVID-19 rapid antigen test will enable medical professionals, emergency workers, and employers to test individuals for antigens that indicate COVID-19 infection. The assay uses a nasal swab sample that can be self-administered, and is designed to complement existing test methods by expanding access to rapid, laboratory-quality results outside of hospital facilities.

“While the global healthcare community has made tremendous progress in COVID-19 testing, there is still significant need for rapid, accurate, and affordable testing that can be conducted outside of laboratory settings,” said David Ludvigson, president and CEO of Nanomix. “By completing this EUA filing, we are one step closer to making our antigen test available to help address this remaining gap.”

Nanomix previously validated and made available a rapid test for IgG and IgM antibodies that also runs on the eLab analyzer, and indicates past exposure to the SARS-CoV-2 virus.

Nanomix developed both the COVID-19 antigen and antibody assays in part with funding from BARDA.

This project has been funded in whole or in part with federal funds from the Department of Health and Human Services; Office of the Assistant Secretary for Preparedness and Response; Biomedical Advanced Research and Development Authority, Division of Research Innovation and Ventures under Contract No. 75A50120C00060.

A New Tool to Investigate Bacteria Behind Hospital Infections

Researchers from the Antimicrobial Resistance (AMR) Interdisciplinary Research Group (IRG) at Singapore-MIT Alliance for Research and Technology (SMART), MIT’s research enterprise in Singapore, and Nanyang Technological University (NTU) have developed a tool using CRISPRi technology that can help understand and prevent biofilm development, drug resistance, and other physiological behaviors of bacteria such as Enterococcus faecalis.

E. faecalis, which is found in the human gut, is one of the most prevalent causes of hospital-associated infections and can lead to a variety of multidrug-resistant, life-threatening infections including bacteremia (bloodstream infection), endocarditis (infection of the heart), catheter-associated urinary tract infection, and wound infections.

However, current methods for understanding and preventing E. faecalis biofilm formation and development are labor-intensive and time-consuming. The SMART AMR research team designed an easily modifiable genetic technique that allows rapid and efficient silencing of bacteria genes to prevent infections.

In a paper published in the journal mBio, the researchers explain the scalable dual-vector nisin-inducible CRISPRi system, which can identify genes that allow bacteria like E. faecalis to form biofilms, cause infections, acquire antibiotic resistance, and evade the host immune system. The team combined CRISPRi technology with rapid DNA assembly under controllable promoters, which enables rapid silencing of single or multiple genes, to investigate nearly any aspect of enterococcal biology.

“Infections caused by E. faecalis are usually antibiotic-tolerant and more difficult to treat, rendering them a significant public health threat,” says Irina Afonina, postdoc at SMART AMR and lead author of the paper. “Identifying the genes that are involved in these bacterial processes can help us discover new drug targets or propose antimicrobial strategies to effectively treat such infections and overcome antimicrobial resistance.”

The team believes their new tool will be valuable in rapid and efficient investigation of a wide range of aspects of enterococcal biology and pathogenesis, host-bacterium interactions, and interspecies communication. The method can be scaled up to simultaneously silence multiple bacterial genes or perform full-genome studies.

“Bacterial biofilms are clusters of bacteria that are enclosed in a protective, self-produced matrix,” says SMART AMR principal investigator and NTU Associate Professor Kimberly Kline, also the corresponding author of the paper. “The system we designed enables us to easily interrogate various stages during the biofilm developmental cycle of E. faecalis. By selectively silencing certain genes in preformed, mature biofilms, we can erode the biofilm and force it to disperse.”

The scalable CRISPRi system uses high-throughput screens that can allow for rapid identification of gene combinations to be simultaneously targeted for novel and efficient antimicrobial combinatorial therapies.

The idea behind SMART’s inducible CRISPRi system was conceived by Kline and SMART AMR principal investigator Professor Timothy Lu, an associate professor in the MIT departments of Electrical Engineering and Computer Science and Biological Engineering, while Afonina developed and delivered the genetic tool.

The research is carried out by SMART and supported by the National Research Foundation (NRF) Singapore under its Campus for Research Excellence And Technological Enterprise (CREATE) program.

SMART was established by MIT in partnership with the NRF Singapore in 2007. SMART is the first entity in CREATE developed by NRF. SMART serves as an intellectual and innovation hub for research interactions between MIT and Singapore, undertaking cutting-edge research projects in areas of interest to both Singapore and MIT. SMART currently comprises an Innovation Center and five IRGs: AMR, Critical Analytics for Manufacturing Personalized-Medicine, Disruptive and Sustainable Technologies for Agricultural Precision, Future Urban Mobility, and Low Energy Electronic Systems.

The AMR IRG is a translational research and entrepreneurship program that tackles the growing threat of antimicrobial resistance. By leveraging talent and convergent technologies across Singapore and MIT, they tackle AMR head-on by developing multiple innovative and disruptive approaches to identify, respond to, and treat drug-resistant microbial infections. Through strong scientific and clinical collaborations, they provide transformative, holistic solutions for Singapore and the world.