Abstract
Since 2020, humanity has been facing the COVID-19 pandemic, a respiratory disease caused by the SARS-CoV-2. The world’s response to pandemic went through the development of diagnostics, vaccines and medicines. Regarding diagnostics, an enormous challenge was faced due to shortage of materials to collect and process the samples, and to perform reliable mass diagnosis by RT-qPCR. In particular, time-consuming and high cost of nucleic acid extraction procedures have hampered the diagnosis; moreover, several steps in the routine for the preparation of the material makes the extracted sample susceptible to contamination. Here two rapid nucleic acid extraction reagents were compared as extraction procedures for SARS-CoV-2 detection in clinical samples by singleplex and multiplex RT-qPCR analysis, using different transport media, samples with high and low viral load, and different PCR machines. As observed, rapid nucleic acid extraction procedures can be applied for reliable diagnosis using a TaqMan-based assay, over multiple platforms. Ultimately, prompt RNA extraction may reduce costs with reagents and plastics, the chances of contamination, and the overall time to diagnosis by RT-qPCR.
Highlights
COVID-19, caused by SARS-CoV-2, became a global pandemic disease from March of2020, when the World Health Organization (WHO) declared a “public health emergency of international concern” [1]
Biological samples for this study were collected from patients suspected of COVID-19 between August 2020 and August 2021
The original Ct values used for diagnosis are shown in Supplementary Tables S1 and S2, and were obtained using PureLink RNA
Summary
COVID-19, caused by SARS-CoV-2, became a global pandemic disease from March of. 2020, when the World Health Organization (WHO) declared a “public health emergency of international concern” [1]. The first case was reported in 2019 in the city of Wuhan, China, after the emergence of symptoms similar to pneumonia in part of the population [2]. The virus that causes the disease belongs to the Coronaviridae family, which has an RNA genome. There is no defined treatment to eliminate the virus, but vaccines are already available to control the disease progress [5–7]; the transmission of SARS-CoV-2 is very fast, and the rate of vaccination is usually slow in several countries, allowing new variants of the virus to emerge around the world [8]
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