Point-of-care diagnostics of COVID-19 in wastewater

Despite major coronavirus immunisation programmes across the world, rigorous public health surveillance and fast diagnostic tests are regarded as the most effective ways to reduce COVID-19. The reverse transcriptase quantitative polymerase chain reaction is the gold standard for diagnosing COVID-19 (RT-qPCR). Droplet digital RT-PCR (RT-ddPCR) provides an appealing platform for quantifying SARS-CoV-2 RNA. High sensitivity and specificity, the necessity for highly skilled workers, and the need for special facilities and expensive apparatus all limit its adoption, particularly in poor nations.

Reverse transcription loop-mediated isothermal amplification (RT-LAMP) is an isothermal nucleic acid amplification technology that is commonly utilised for SARS-CoV-2 detection in clinical samples. Because of its inherent benefits such as sensitivity, speed, the absence of a heat cycler, and resistance to sample inhibitors, RT-LAMP is a prospective alternative to RT-qPCR. LAMP takes less than an hour to amplify the pathogen's genetic material and requires a set of four to six primers, ensuring great specificity. LAMP amplification products can be validated using several approaches such as changes in fluorescence utilising intercalating dyes, DNA probes with gold nanoparticles, changes in turbidity generated by magnesium pyrophosphate precipitate, pH indicators, or gel electrophoresis followed by UV detection. For quick and easy detection, the most often used approach is based on color change of a colorimetric master mix including a visible pH indicator.

Although aerosol/droplet inhalation and person-to-person contact are the primary routes of SARS-CoV-2 transmission, current evidence indicates that viral RNA is detected in wastewater, highlighting the need to better understand wastewater as a potential source of epidemiological data and human health risks that can be used as an early warning system. SARS-CoV-2 can generate asymptomatic or pauci-symptomatic infections, which complicates determining the true extent of SARS-CoV-2 circulation in a population. Meanwhile, wastewater surveillance can provide a less biassed means of predicting the spread of illness in diverse locations, particularly in underdeveloped nations where clinical diagnostic tools are scarce and restricted.

References

Donia, Ahmed, et al. "Integration of RT-LAMP and Microfluidic Technology for Detection of SARS-CoV-2 in Wastewater as an Advanced Point-of-care Platform." Food and environmental virology (2022): 1-10.

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