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