Perez Vinícius Pietta, Pessoa Wallace Felipe Blohem, Galvão Bruno Henrique Andrade, Sousa Eduardo Sergio Soares, Dejani Naiara Naiana, Campana Eloiza Helena, Cavalcanti Marilia Gabriela Dos Santos, Cantarelli Vlademir Vicente
Departamento de Fisiologia e Patologia, Centro de Ciências da Saúde, Universidade Federal da Paraíba, Campus I - Cidade Universitária s/n, João Pessoa, PB 58051-900, Brazil.
LaBiMol, Centro de Ciências Médicas, Universidade Federal da Paraíba, João Pessoa, Brazil.
J Clin Virol Plus. 2021 Sep;1(3):100032. doi: 10.1016/j.jcvp.2021.100032. Epub 2021 Jul 3.
The efficiency of isolation and purification of the viral genome is a critical step to the accuracy and reliability of RT-qPCR to detect SARS-CoV-2. However, COVID-19 testing laboratories were overwhelmed by a surge in diagnostic demand that affected supply chains especially in low and middle-income facilities.
Thus, this study compares the performance of alternative methods to extraction and purification of viral RNA in samples of patients diagnosed with COVID-19.
Nasopharyngeal swabs were submitted to three protocols and three commercial methods; viral genome was detected using the primer-probe (N1 and N2) described by CDC and viral load of samples were determined.
The protocols resulted in detection of virus in 82.4 to 86.3% of samples and commercial methods in 94.1 to 98%. The disagreement results were observed in samples with low viral load or below the estimated limit of detection of RT-qPCR.
The simplified methods proposed might be less reliable for patients with low viral load and alternative commercial methods showed comparable performance.
病毒基因组的分离和纯化效率是RT-qPCR检测严重急性呼吸综合征冠状病毒2（SARS-CoV-2）准确性和可靠性的关键步骤。然而，新型冠状病毒肺炎（COVID-19）检测实验室因诊断需求激增而不堪重负，这影响了供应链，尤其是在低收入和中等收入机构。
因此，本研究比较了COVID-19确诊患者样本中病毒RNA提取和纯化替代方法的性能。
鼻咽拭子采用三种方案和三种商业方法进行处理；使用美国疾病控制与预防中心（CDC）描述的引物-探针（N1和N2）检测病毒基因组，并测定样本的病毒载量。
三种方案在82.4%至86.3%的样本中检测到病毒，商业方法在94.1%至98%的样本中检测到病毒。在病毒载量低或低于RT-qPCR估计检测限的样本中观察到结果不一致。
对于病毒载量低的患者，所提出的简化方法可能不太可靠，而替代商业方法表现出相当的性能。
