Comparative performance of SARS-CoV-2 real-time PCR diagnostic assays on samples from Lagos, Nigeria

• DOI: 10.1371/journal.pone.0246637
• Publication Date: 2021-02-05T03:52:00Z
• Authors: Chika Kingsley On...; Azuka Patrick Okw...; Olumuyiwa B. Salu; Joseph O. Shaibu; Nnaemeka Ndodo; Samuel O. Amoo; Leona C. Okoli; Fehintola A. Ige; Rahaman A. Ahmed; Munir Akinwale Ba...; Judith O. Sokei; Bamidele Paul Mutiu; James Ayorinde; Babatunde Akeem Saka; Celestina Obiekea; Nwando Mba; Richard A. Adegbola; Sunday Omilabu; Chikwe Ihekweazu; Babatunde Lawal S...; Rosemary Audu
• Keywords: 0301 basic medicine; SARS-CoV-2; Science; Q; R; COVID-19; Nigeria; Sensitivity and Specificity; 3. Good health; 03 medical and health sciences; COVID-19 Nucleic Acid Testing; Medicine; Humans; RNA, Viral; Research Article; Retrospective Studies

A key element in containing the spread of the SARS-CoV-2 infection is quality diagnostics which is affected by several factors. We now report the comparative performance of five real-time diagnostic assays. Nasopharyngeal swab samples were obtained from persons seeking a diagnosis for SARS-CoV-2 infection in Lagos, Nigeria. The comparison was performed on the same negative, low, and high-positive sample set, with viral RNA extracted using the Qiagen Viral RNA Kit. All five assays are one-step reverse transcriptase real-time PCR assays. Testing was done according to each assay’s manufacturer instructions for use using real-time PCR platforms. 63 samples were tested using the five qPCR assays, comprising of 15 negative samples, 15 positive samples (Ct = 16–30; one Ct = 35), and 33 samples with Tib MolBiol E-gene Ct value ranging from 36–41. All assays detected all high positive samples correctly. Three assays correctly identified all negative samples while two assays each failed to correctly identify one different negative sample. The consistent detection of positive samples at different Ct/Cq values gives an indication of when to repeat testing and/or establish more stringent in-house cut-off value. The varied performance of different diagnostic assays, mostly with emergency use approvals, for a novel virus is expected. Comparative assays’ performance reported may guide laboratories to determine both their repeat testing Ct/Cq range and/or cut-off value.