Evaluation of Currently Available Molecular Assays and Performance of Sampling Approaches for Detection of Sars-Cov-2 RNA

Abstract

OBJECTIVES This study aims to identify the essential characteristics of diagnostic tests for SARS-CoV-2 and to discuss the limitations of currently available tests and their impact on the test selection process. METHODOLOGY The current study was conducted at Mardan Medical Complex (MMC). One hundred nasopharyngeal-positive samples were collected from February to March 2021. Oropharyngeal swab OPS, sputum, and blood samples were collected from the participants to detect SARS-CoV-2 RNA. RNA extraction of SARS-CoV-2 was done using a BigFish auto extractor. A Qiagen Thermal Cycler was used for genome amplification. Five different molecular assays, namely COVSIGN (N gene) Spain, BGI (ORF1ab gene) China, Maccura(ORF1ab, E and N gene) China, R-GENE (RdRp and N genes) France and Genuru (N gene, S gene and ORF ab/1) were used. RESULTS100 % positivity was recorded in the sputum of all individuals, followed by 91 % OPS and 21% blood. The highest positivity rate for different genes was observed. ROC (Receiver operating characteristic curve) was developed through SPPS version 26.00 to compare the sensitivity and specificity. CONCLUSION By comparing the results of different diagnostic kits, it was found that BGI and Maccura are the most sensitive and specific for diagnostic purposes against COVID-19.