For those who have been to doctors and clinics during the upcoming flu season, the most annoying concern is whether their symptoms indicate a bug in the cultivars that are prevalent in the workplace, or COVID-19 infections. Is it showing the possibility? When researchers scramble to treat this virus and its unavoidable mutations and mutations, one of the biggest challenges is to ensure that proper treatment protocols are implemented, and potentially dangerous. To reduce sex, the cause of the disease was to be diagnosed quickly and accurately. complications.
At the University of Florida, led by Dr. Z. Hugh Fan, a professor of mechanical and aerospace engineering at the Herbert Wertheim Institute of Technology, and Dr. John Rednicky, a research professor of the Department of Environmental Studies at the University of Public Health and Health Professions. Global Health, an interdisciplinary team, has developed a groundbreaking game Diagnostic test For SARS-CoV-2, which is fast, reliable, low cost, and can distinguish between COVID-19 and influenza.
Dr. Huang and his team have demonstrated the ability to detect two viruses in 50 minutes without the use of bulky or sophisticated laboratory equipment or power supplies. Detection sensitivity This is equivalent to the Gold Standard RT-PCR (Reverse Transcription Polymerase Chain Reaction) assay, a test recommended by the World Health Organization and the US Center for Disease Control and Prevention. This effort is supported by the National Institutes of Health.
“From a patient care perspective, this innovative sample-to-answer platform is a significant upgrade from existing RT-PCR standards at several levels,” said Dr. Fan. “When it comes to diagnostics, it takes less than an hour at the test spot compared to a day or two from an offsite test lab.”
A device designed at the University of Florida was recently published for peer review. ACS sensor, The flagship journal of the American Chemical Society with a focus on sensor science.
RT-LAMP (loop-mediated isothermal amplification) is an assay used to detect viral RNA. The RT-LAMP methodology was based on the mechanism behind automated cycling strand substitution DNA synthesis. The polymerase reacts and the polymerase has high chain substitution activity. There are also two pairs of primers used, one is the pair of internal primers and the other is the pair of external primers. These primers are specially designed for the reaction. RT-LAMP achieves higher specificity due to the target sequence. Unlike other techniques, RT-LAMP initially uses six independent sequences to recognize the target sequence and give it high specificity. Primer recognition of the target genome results in a strong colorimetric response, enabling detection without highly specialized or costly equipment.
“RT-LAMP technology has many advantages over standard RT-PCR,” says Dr. Lednicky. “For use in users and clinics in countries where reagent costs are lower, results are obtained more quickly, RT-LAMP equipment needs are lower, and resources are scarce compared to comparable RT-PCR reactions. Also, the RT-LAMP reaction packaged in a kit format requires very little storage space and can be freeze-dried, reducing the need for storage in the freezer. “
Depending on the test, the sample (cotton swab, urine, saliva, blood) is taken and the measured amount is transferred to the first chamber of the sample post-treatment device. The specimen is then exposed to a specific liquid that extracts DNA or RNA from the pathogen, and the extracted material is purified and collected in a removable filter unit. A microchip-controlled “coffee mug” with RT-LAMP chemicals added to the removable unit, the chamber containing the RT-LAMP liquid sealed with tape, the last unit removed, and preheated to a specific temperature. Can be put in. .. After the selected incubation period, the results are determined and evidenced by a color change that is either visually detected or also detectable / measurable by its fluorescence level.
The disposable unit in disposable format allows nurses, doctors, etc. to use this device without the need for advanced training. Whether positive or negative, the results can be judged visually or using a simple technique such as a flashlight. Results can be recorded by capturing the image using an iPhone or similar mobile device with a camera and sending it to a remote site to review the data.
“In conclusion, this device enables faster analysis of genetic material than traditional RT-PCR and has been successfully used to detect the SARS-CoV-2 virus,” said Dr. Lednicky. “Simple, inexpensive, and does not require advanced technical skills or expensive equipment. Most importantly, it is extremely sensitive, sensitive to detection, and in some cases 10 times more than standard RT-PCR assays. It’s about getting sensitivity. “
Currently, simultaneous testing of both SARS-CoV-2 and influenza virus is not available at the Point of Care (POC), but variations are available in the test lab and may be considered a “CLIA exempt” device. Includes FDA-approved home tests. “This device is Environmental studies It can be pasted / coupled with (SARS-CoV-2 in air, sewage, or other matrices), and other detectors such as air samplers, “says Dr. Fan.
The development of this LAMP-based device follows the progressive progress made by UF researchers in the early detection and identification of dangerous pathogens. Three years ago, funded by Florida, Dr. Fan and Dr. Rednicky led a research team that developed a fast, cost-effective, miniaturized device for detecting mosquito-borne Zika virus. did. To 80% of infected people. The Point of Care testing platform helps screen asymptomatic patients in the field, reduce misdiagnosis, and prevent further infections.
Last year, an engineer in the Department of Environmental Engineering at the Department of Sustainable Infrastructure and Environmental Engineering (ESSIE), led by Dr. Chang-Yu Wu, developed a new viral aerosol sampler that efficiently collects airborne viruses (including COVID-19). .. Information for healthcare providers to identify risk areas and identify measures for improvement. With the integration of the new sampler and RT-LAMP device, the virus goes beyond the test area to reach public areas (airports, subways, government buildings, shopping centers, etc.), providing early warning of aerial pathogenic viruses. Can stop the spread of the virus.
Carlos Manzanas et al, valve-compatible sample preparation device with isothermal amplification for multiple virus detection at the point of care, ACS sensor (2021). DOI: 10.1021 / acssensors.1c01718
University of Florida
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