Conventional reverse transcription-polymerase chain reaction assay as an alternative, low-cost, and reliable method for the detection of COVID-۱۹

Background: Severe acute respiratory syndrome coronavirus ۲ (SARS-CoV-۲) is a highly contagious infection causing a large number of deaths in susceptible individuals throughout the world. Objectives: In this study, a low-cost, sensitive, and easy-to-perform conventional Polymerase chain reaction (PCR)-based RNA detection method was evaluated to diagnose the infection that was feasible at a laboratory with minimal molecular infrastructure. Methods: From ۴ July to ۳۱ August ۲۰۲۰, a total of ۲۷۷ nasopharyngeal/oropharyngeal swab samples consisting of ۷۲ samples from hospitalized patients with a severe respiratory infection and ۲۰۵ suspected patients into four different age groups of under ۲۰ years (n=۸), ۲۰-۳۹ years (n=۹۰), ۴۰-۵۹ years (n=۹۳), and over ۶۰ years (n=۸۶) in Isfahan, Iran, were tested using the real-time reverse transcription polymerase chain reaction (rRT-PCR), and conventional PCR assays. Results: Overall, ۴۰ cases out of ۷۲ hospitalized patients were aged more than ۶۰ years, and the majority of the study population were male (۵۴.۱%, n=۱۵۰). Out of ۱۶۰ clinical samples tested by RT-PCR with the E gene, the sensitivity and specificity of the conventional PCR method were determined at ۱۰۰%. Furthermore, out of ۱۱۷ clinical samples evaluated by the probe-based RT-PCR with the N gene, ۷۴.۴% of the samples were positive. Moreover, the duplex PCR method using the N gene and RNase P as an internal control reference gene showed that ۶۸.۴% of the samples were positive. Therefore, the tested PCRs could detect positive samples with a sensitivity of ۹۲.۵۵% and a specificity of ۱۰۰%. Conclusion: According to the results, this method is a simple, inexpensive, and valuable alternative as well as a suitable procedure for the laboratory diagnosis of SARS-CoV-۲ infection.

Keywords:

COVID-۱۹ , Reverse transcription-polymerase chain reaction , Diagnosis , COVID-۱۹ , Reverse transcription-polymerase chain reaction , Diagnosis