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.