Rapid, Reliable and Robust Approach for Extraction-Free RT-PCR based Detection of SARS-CoV-2in Clinical Setting to Expedite Large Scale Screening

Abhilasha Dubey, Sanjay Upadhyay, Manjeet Mehta

Abstract: Rapid, reliable and robust method for the detection of SARS-CoV-2 is an indispensable need for diagnostics. The development of diagnostic method will aid to address further waves of the pandemic potentially with rapid surveillance of disease; and to allay the fears. To meet this challenge, we have developed a rapid RT-qPCR method for the detection of 3 target genes or confirmatory genes in less than 30 minutes. The assay showed 100% sensitivity and 100% specificity when tested on 120 samples. We compared a conventional extraction based method with extraction-free method, and then further reduced the run time of extraction free method. Additionally, we have validated our rapid RT-qPCR method for the assessment of pooled sample. We here by propose a most reliable approach for the mass screening of samples with ease of operation at low cost. Finally we designed a single tube analysis method which provides qualitative as well as quantitative results in minimum time.

Keywords: SARS-CoV-2, COVID-19, RT-PCR, heat treatment, nasopharyngeal swab samples, Extraction free method, Pooling

How to Cite?: Abhilasha Dubey, Sanjay Upadhyay, Manjeet Mehta, "Rapid, Reliable and Robust Approach for Extraction-Free RT-PCR based Detection of SARS-CoV-2in Clinical Setting to Expedite Large Scale Screening", Volume 10 Issue 11, November 2021, International Journal of Science and Research (IJSR), Pages: 1378-1385, https://www.ijsr.net/getabstract.php?paperid=SR211126224931, DOI: https://dx.doi.org/10.21275/SR211126224931