Abstract INTRODUCTION: The main laboratory test for the diagnosis of coronavirus disease 2019 (COVID-19) is the reverse transcription real-time polymerase chain reaction (RT-qPCR). However, RT-qPCR is expensive because of the number of tests required. This study aimed to evaluate an alternative to the RT-qPCR approach for the detection of sudden acute respiratory syndrome coronavirus 2 (SARS-CoV-2) that is half of the total volume currently recommended by the US Centers for Disease Control and Prevention. METHODS: The analytical limit of detection (LoD) and the reaction efficiency using half volumes of the RT-qPCR assay were evaluated for the N1 and N2 regions using a synthetic control RNA. A panel of 76 SARS-CoV-2-positive and 26 SARS-CoV-2-negative clinical samples was evaluated to establish clinical sensitivity and specificity. RESULTS: The RT-qPCR assay efficiency was 105% for the half and standard reactions considering the N2 target and 84% (standard) and 101% (half) for N1. The RT-qPCR half-reaction LoD for N1 and N2 were 20 and 80 copies/µL, respectively. The clinical sensitivity and specificity were 100%. The half reaction presented a decrease of up to 5.5 cycle thresholds compared with standard RT-qPCR. CONCLUSIONS: The use of the RT-qPCR half-reaction proved feasible and economic for the detection of SARS-CoV-2 RNA.
【저자키워드】 COVID-19, SARS-CoV-2, Coronavirus disease 2019, coronavirus, RT-qPCR half-reaction, 【초록키워드】 coronavirus disease, Coronavirus disease 2019, coronavirus, Diagnosis, prevention, Laboratory, RNA, polymerase chain reaction, clinical samples, specificity, RT-qPCR, Region, cycle threshold, disease control, limit of detection, Control, reverse transcription, SARS-CoV-2 RNA, respiratory, Efficiency, clinical sensitivity, Laboratory test, Chain Reaction, Volume, acute respiratory syndrome, alternative, reaction, reverse transcription real-time polymerase chain reaction, center, N1 and N2, approach, decrease, polymerase chain, clinical sample, evaluate, evaluated, required, feasible,