Evaluating the efficiency of ELISA, monoplex and multiplex probe‐based real‐time reverse‐transcription PCR assays in the detection of SARS‐CoV‐2 (COVID‐19) and influenza A and B viruses: A cross‐sectional study

Background and Aims The current study aimed to evaluate the efficiency of Enzyme‐linked immunosorbent assay (ELISA) assay and monoplex and multiplex real‐time reverse‐transcription PCR (rRT‐PCR) in the detection of severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) and influenza A and B viruses (Flu A and Flu B). Methods The SARS‐CoV‐2 ‐specific IgG and IgM antibodies, as well as, Flu A (H1N1 and H3N2 serotypes) and Flu B virus antibodies were determined by ELISA assay. The one‐step qRT‐PCR method was used to detect the SARS‐CoV‐2 in nasopharyngeal swab samples. Furthermore, the presence of Flu A and B viruses was evaluated using probe‐based RT‐PCR. Simultaneous detection of SARS‐CoV‐2, Flu A and B viruses was performed by multiplex rRT‐PCR assay. Results SARS CoV‐2 IgM and IgG antibodies were detected in 33.3% and 58.3% of patients, respectively. In contrast, the SARS CoV‐2 genome was detected in 50% of patients using the one‐step monoplex RT‐PCR assay. Flu A serotypes H1N1 and H3N2 were found in 16.7% and 8.3% of patients. Probe‐based RT‐PCR revealed that 39.3% of patients were positive for the Flu A virus. Multiplex rRT‐PCR detect the SARS‐CoV‐2, Flu A, and Flu B in 50%, 39.3%, and 19% of samples, respectively. The sensitivity and specificity of multiplex rRT‐PCR assay in comparison to monoplex RT‐PCR were 100% and 55%, respectively. Coinfection with SARS‐CoV‐2, Flu A, and Flu B viruses was found in 9.5% of patients. Conclusion Multiplex rRT‐PCR can be used as a repaid, cost‐effective and suitable tool for molecular surveillance of SARS‐CoV‐2 and Flu A/B viruses.