Research Unit of Systems Microbiology, Faculty of Medicine, Chulalongkorn University, Bangkok, 10330, Thailand.
Department of Biochemistry, Faculty of Medicine, Chulalongkorn University, Bangkok, 10330, Thailand.
J Virol Methods. 2021 Apr;290:114092. doi: 10.1016/j.jviromet.2021.114092. Epub 2021 Feb 1.
COVID-19 pandemic caused by SARS-CoV-2 infection continue to cause the morbidity and mortality in many countries. Limitations of the gold standard qRT-PCR for diagnosis of this infection includes need for expensive equipment, specialized molecular laboratory, and experienced staff. Currently, CRISPR-based diagnostic method was approved by the U.S. FDA for rapid detection. Several studies developed SARS-CoV-2 detection based on CRISPR-Cas12a platform; however, the validations with RNA extracted from clinical specimens were limited. Therefore, this study evaluated the clinical performance of previously described CRISPR-Cas12a based diagnostic assays for SARS-CoV-2. According to the results, the CRISPR-Cas12a assays on N1 and S genes provided diagnostic accuracy (≥ 95 %) comparable to the qRT-PCR results. The assays with E, N2 and S genes yielded acceptable sensitivity of detection (≥ 95 %) whereas N1 and S genes provided outstanding specificity of detection (100 %). Preferably, multiple target genes should be detected by using CRISPR-Cas12a to ensure the most effective SARS-CoV-2 detection. Therefore, the N1 and S genes would be attractive target genes for SARS-CoV-2 detection based on CRISPR-Cas12a.
由 SARS-CoV-2 感染引起的 COVID-19 大流行继续在许多国家造成发病率和死亡率。金标准 qRT-PCR 诊断这种感染的局限性包括需要昂贵的设备、专门的分子实验室和有经验的人员。目前，基于 CRISPR 的诊断方法已获得美国 FDA 批准，可用于快速检测。有几项研究基于 CRISPR-Cas12a 平台开发了 SARS-CoV-2 检测方法；然而，从临床标本中提取 RNA 的验证有限。因此，本研究评估了先前描述的基于 CRISPR-Cas12a 的 SARS-CoV-2 诊断检测的临床性能。结果表明，N1 和 S 基因的 CRISPR-Cas12a 检测提供了与 qRT-PCR 结果相当的诊断准确性（≥95%）。具有 E、N2 和 S 基因的检测方法具有可接受的检测灵敏度（≥95%），而 N1 和 S 基因提供了出色的检测特异性（100%）。最好使用 CRISPR-Cas12a 检测多个靶基因，以确保最有效的 SARS-CoV-2 检测。因此，N1 和 S 基因将是基于 CRISPR-Cas12a 的 SARS-CoV-2 检测的有吸引力的靶基因。
