Background The emergence of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) pandemic is posing a great threat to global health and economy. Due to the lack of broad diagnostic setup, consistent reagent supply lines, and access to laboratory instruments and equipment, it is undoubtedly an enormous burden for developing countries to face the crisis. Objectives To develop a cost-effective, reliable and sensitive multiplex assay for SARS-CoV-2 screening which would expand the testing capacities of a developing and low-income country like Bangladesh. Study design Initially a singleplex and then a multiplex real-time reverse-transcriptase PCR assays were developed targeting 2 nucleocapsid genes of SARS-CoV-2, and the human RNase P gene as an internal control using laboratory-made mastermixes. Three sets of primer- probes were designed for each of the target genes and one set was optimized for the final reaction set-up. Limit of detection, cross-reactivity and reproducibility were checked in order to assess the sensitivity and specificity of the assays, and validation was done using clinical specimens. Results Clinical evaluation of the new assays using 240 nasopharyngeal swabs showed 100 % sensitivity, specificity, and accuracy in detecting SARS-CoV-2 infection in human. Equal efficiency and concordant results were observed between the singleplex and multiplex approaches. Notably, the kit was able to detect SARS-CoV-2 RNA at very low concentration upto 5 copies/reaction. Conclusion This is the first locally developed multiplex rRT-PCR kit in Bangladesh providing rapid and low-cost screening of COVID-19 which would be valuable for infection prevention and clinical management in the perspective of Bangladesh.
【저자키워드】 SARS-CoV-2, developing country, Singleplex rRT-PCR, Multiplex rRT-PCR, Cost effective assay, In-house kit, 【초록키워드】 COVID-19, pandemic, equipment, SARS-COV-2 infection, diagnostic, Laboratory, cross-reactivity, Nasopharyngeal swab, sensitivity, specificity, rRT-PCR, Health, Multiplex assay, Accuracy, Sensitivity and specificity, Internal control, clinical, Infection prevention, Clinical management, SARS-CoV-2 RNA, multiplex, specimens, Nucleocapsid gene, Coronavirus-2, Concentration, Efficiency, target gene, acute respiratory syndrome, approaches, Perspective, Equal, Final, primer, RNase P, reproducibility, reverse-transcriptase PCR, probe, objective, country, Result, detect, lack, develop, assays, expand, checked, was done, Initially, 【제목키워드】 rRT-PCR, multiplex, development,