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Game-Changing Diagnostic Test for SARS-CoV-2 Rapidly Differentiates Between COVID-19 and Influenza

By LabMedica International staff writers
Posted on 11 Jan 2022
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Image: Assembled Unit (Photo courtesy of University of Florida)
Image: Assembled Unit (Photo courtesy of University of Florida)

An interdisciplinary team of researchers has developed a game-changing diagnostic test for SARS-CoV-2 that is fast, reliable, low-cost and capable of differentiating between COVID-19 and influenza.

The researchers at the University of Florida (Gainesville, FL, USA) who developed the test also demonstrated its ability to detect two viruses in 50 minutes without using bulky or sophisticated laboratory equipment or power supply, and with detection sensitivity that is on par with that of the gold-standard RT-PCR (reverse transcription polymerase chain reaction) assay, which is the test recommended by the World Health Organization and the Centers for Disease Control and Prevention.

For those who have made a trip to a doctor or clinic during the oncoming flu season, the most nagging concern is whether their symptoms are indicative of the garden-variety bug that is making the rounds in the workplace or a potential COVID-19 infection. As researchers scramble to treat this virus - and its inevitable variants and mutations - one of the biggest challenges has been quickly and accurately diagnosing the source of the illness so that the proper treatment protocols are administered, and in time to decrease the odds of potentially dangerous complications.

RT-LAMP (loop-mediated isothermal amplification) is an assay used for viral RNA detection. The researchers developed a LAMP-based device that may allow clinicians to tell COVID-19 from seasonal flu with a fast turnaround. The methodology for RT-LAMP was based on the mechanism behind auto-cycling strand displacement DNA synthesis. A polymerase carries out the reaction, and the polymerase has high strand displacement activity. There are also two pairs of primers used, one pair of inner and another of outer primers. These primers are specially designed for the reaction. RT-LAMP achieves high specificity due to the target sequences. Unlike other technologies, RT-LAMP recognizes the target sequence using six independent sequences at the start, giving it high specificity. Primer recognition of the target genome leads to a strong colorimetric reaction, which allows for detection without highly specialized or costly instrumentation.

The specimen - swab, urine, saliva, blood, depending on the test - is taken, transferring a determined amount into the first chamber of the sample work-up device. Sequentially, the specimen is subjected to specific fluids that extract DNA or RNA from the pathogen, purifying the extracted material and collecting it on a detachable filter unit. RT-LAMP chemicals are added into the detachable unit, the chamber with the RT-LAMP fluids is sealed with tape and the final unit is detached and placed into a microchip-controlled “coffee mug,” which is pre-heated to a specific temperature. After a selected incubation period, the results are determined, evidenced by a color change that is visually detected or that can also be detected/measured through its level of fluorescence.

Using disposable-format single-use units, the device can be used by nurses, doctors, and others without the need for advanced training. The results, positive or negative, can be determined visually or with the aid of simple technology like a flashlight. The results can be recorded by using an iPhone or similar camera-equipped mobile device to capture the image, where it is sent to a remote site for confirmation of the data. The simultaneous test of both SARS-CoV-2 and influenza viruses isn’t presently available at point-of-care (POC), but its variations are available in test labs, and would likely be considered as a “CLIA-waived” device, which includes home tests approved by the FDA.

“In patient-care terms, this innovative sample-to-answer platform is an important upgrade from the existing RT-PCR standard on several levels,” said Dr. Z. Hugh Fan, Ph.D., professor at the Herbert Wertheim College of Engineering’s Department of Mechanical and Aerospace Engineering, who led the team. “With regard to diagnosis, the turnaround time is less than an hour at the testing spot compared to one to two days from an offsite testing lab.”

“Bottom line, the device allows for quicker analysis of genetic material than traditional RT-PCR and has been successfully used in the detection of the SARS-CoV-2 virus,” said Dr. John Lednicky, Research Professor at the College of Public Health and Health Professions’ Department of Environmental and Global Health. “It’s simple, cheap, and does not require advanced technical skills or expensive equipment. Most importantly, it’s very sensitive, yielding detection sensitivities, in some cases, 10 times more sensitive than standard RT-PCR assays.”

Related Links:
University of Florida 

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