The ongoing COVID-19 pandemic has created an unprecedented need for rapid diagnostic testing. The World Health Organization (WHO) recommends a standard assay that includes an RNA extraction step from a nasopharyngeal (NP) swab followed by reverse transcription–quantitative polymerase chain reaction (RT-qPCR) to detect the purified SARS-CoV-2 RNA. The current global shortage of RNA extraction kits has caused a severe bottleneck to COVID-19 testing. The goal of this study was to determine whether SARS-CoV-2 RNA could be detected from NP samples via a direct RT-qPCR assay that omits the RNA extraction step altogether. The direct RT-qPCR approach correctly identified 92% of a reference set of blinded NP samples ( n = 155) demonstrated to be positive for SARS-CoV-2 RNA by traditional clinical diagnostic RT-qPCR that included an RNA extraction. Importantly, the direct method had sufficient sensitivity to reliably detect those patients with viral loads that correlate with the presence of infectious virus. Thus, this strategy has the potential to ease supply choke points to substantially expand COVID-19 testing and screening capacity and should be applicable throughout the world. A major bottleneck to RT-PCR-based COVID-19 testing is the RNA extraction step. This study shows that a “direct RT-PCR” approach that omits the RNA extraction step can reliably detect SARS-CoV-2 RNA in nasopharyngeal swabs from individuals most likely to be infectious.
【초록키워드】 COVID-19 pandemic, diagnostic, RNA extraction, Nasopharyngeal swab, polymerase chain reaction, sensitivity, RT-qPCR, COVID-19 testing, Viral, Viral load, nasopharyngeal swabs, nasopharyngeal, Patient, diagnostic testing, WHO, SARS-CoV-2 RNA, Swab, infectious, Infectious virus, followed by, Health Organization, be positive, Chain Reaction, choke points, direct method, World Health Organization, individual, viral loads, RNA extraction kits, positive, approach, polymerase chain, detect, caused, blinded, include, determine, demonstrated, purified, expand, RNA extraction kit, 【제목키워드】 Nasopharyngeal swab, nasopharyngeal swabs, Patient, Direct,