Real-time reverse transcription-polymerase chain reaction (RT-qPCR) is considered the "gold standard" for the direct diagnosis of SARS-CoV-2 infections. However, routine diagnosis by RT-qPCR is a limitation for many laboratories, mainly due to the infrastructure and/or disproportionate relationship between demand and supply of inputs. In this context, and to increase the diagnostic coverage of SARS-CoV-2 infections, we describe an alternative, sensitive and specific one-step end-point RT-PCR for the detection of the SARS-CoV-2 E gene. The performance of the RT-PCR was evaluated in 43 clinical samples, of which 10 and 33 were previously identified as negative and positive, respectively, by RT-qPCR. Among the positive samples, 15 and 18 were from asymptomatic and symptomatic individuals, respectively. Here, 32/33 of the positive samples in the RT-qPCR, including from asymptomatic individuals, were found positive in the RT-PCR (Ct 15.94–34.92). The analytical sensitivity of the assay was about 7.15–9 copies of vRNA/μL, and nonspecific amplifications were not observed in SARS-CoV-2 negative samples. Importantly, the RT-PCR reactions were performed in a 10 μL final volume. Finally, considering specificity, analytical sensitivity and cost reduction, we believe that the RT-PCR platform described here may be a viable option for the diagnostic of SARS-CoV-2 infections in laboratories in which RT-qPCR is not available.

实时逆转录聚合酶链反应（RT-qPCR）被认为是直接诊断 SARS-CoV-2 感染的“金标准”。然而，RT-qPCR 的常规诊断对许多实验室来说是一种限制，主要是由于基础设施和/或投入品的供需关系不成比例。在此背景下，为了提高 SARS-CoV-2 感染的诊断覆盖率，我们描述了一种替代的、灵敏且特异的一步终点 RT-PCR，用于检测 SARS-CoV-2 E 基因。RT-PCR 的性能在 43 个临床样本中进行了评估，其中 10 个和 33 个先前通过 RT-qPCR 分别鉴定为阴性和阳性。阳性样本中，无症状者和有症状者分别为15例和18例。在此，RT-qPCR 中 32/33 的阳性样本（包括来自无症状个体）在 RT-PCR 中被发现呈阳性（Ct 15.94–34.92）。该测定的分析灵敏度约为 7.15-9 个 vRNA 拷贝/μL，并且在 SARS-CoV-2 阴性样本中未观察到非特异性扩增。重要的是，RT-PCR 反应的最终体积为 10 μL。最后，考虑到特异性、分析灵敏度和成本降低，我们认为此处描述的 RT-PCR 平台可能是在无法使用 RT-qPCR 的实验室中诊断 SARS-CoV-2 感染的可行选择。