Conventional Reverse Transcription-Polymerase Chain Reaction Assay as an Alternative, Low-Cost, and Reliable Method for the Detection of Coronavirus Disease 2019

Background: sars-cov-2 disease is a highly contagious infection causing a large number of deaths in susceptible individuals throughout the world. 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, which was feasible at a laboratory with minimal molecular infrastructure.methods: from 4 july to 31 august 2020, a total of 277 nasopharyngeal/oropharyngeal swab samples consisting of 72 samples from hospitalized patients with a severe respiratory infection and 205 suspected patients in isfahan, iran, were tested using probe-based rtrt-pcr and conventional pcr assays.results: a total of 160 clinical samples were tested by rtrt-pcr using the e gene. the sensitivity and specificity of the conventional pcr method were determined to be 100%. furthermore, out of 117 clinical samples evaluated by the probe-based rt-pcr using the n gene, 74.4% 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 68.4% of the samples were positive. therefore, the tested pcrs could detect positive samples with a sensitivity of 92.55% and a specificity of 100%.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-2 infection.