Performance Evaluation of Real Time Polymerase Reaction Assay (SCODA) for Detection of SARS-CoV-2

Abstract

In order to robustly detect and quantify gene expression from small amounts of RNA, amplification of the gene transcript is necessary. Real Time PCR is useful for detection and quantification of genetic constitution of pathogens. This technique amplifies a tiny DNA target million or billion times in such a way that it can be easily studied by scientists. Availability of highly sensitive and specific assay for the detection of SARS-Cov-2 and easy accessibility of such was necessary for early diagnosis and effective management of COVID-19 infection. The aim of this study was to evaluate the performance characteristic of SCODA. Validation of SCODA was performed using synthesized standards and clinical samples previously tested using a commercially approved COVID-19 RT-qPCR detection kit (LifeRiver). The assay showed a linearity of 98.2% on the ORF1ab target and 99.8% on the N-gene target. The sensitivity and specificity were both 100%. Analysis for the LoD95 produced 74.04 (CI: 25 - 1000) cp/μl on ORF1ab gene and 1.119 (CI: 1 - 1) cp/μl on N-gene target with a precision of CV ≤ 3%. SCODA showed high comparable performance in comparison with LifeRiver and other commercial COVID-19 RT-qPCR test kits.