Laboratory of Virology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, MT 59840, USA.
Department of Pathology, Microbiology, & Immunology, School of Veterinary Medicine, University of California, Davis, Davis, CA 95616, USA.
Viruses. 2021 Apr 1;13(4):599. doi: 10.3390/v13040599.
Coronavirus (CoV) spillover events from wildlife reservoirs can result in mild to severe human respiratory illness. These spillover events underlie the importance of detecting known and novel CoVs circulating in reservoir host species and determining CoV prevalence and distribution, allowing improved prediction of spillover events or where a human-reservoir interface should be closely monitored. To increase the likelihood of detecting all circulating genera and strains, we have modified primers published by Watanabe et al. in 2010 to generate a semi-nested pan-CoV PCR assay. Representatives from the four coronavirus genera (α-CoVs, β-CoVs, γ-CoVs and δ-CoVs) were tested and all of the in-house CoVs were detected using this assay. After comparing both assays, we found that the updated assay reliably detected viruses in all genera of CoVs with high sensitivity, whereas the sensitivity of the original assay was lower. Our updated PCR assay is an important tool to detect, monitor and track CoVs to enhance viral surveillance in reservoir hosts.
冠状病毒（CoV）从野生动物宿主溢出到人类会引起从轻微到严重的呼吸道疾病。这些溢出事件突显了检测在宿主物种中循环的已知和新型 CoV 的重要性，并确定 CoV 的流行率和分布，从而可以更好地预测溢出事件或需要密切监测的人与宿主之间的界面。为了提高检测所有循环属和菌株的可能性，我们修改了 Watanabe 等人于 2010 年发表的引物，以生成半巢式泛 CoV PCR 检测法。使用该检测法测试了来自四个冠状病毒属（α-CoVs、β-CoVs、γ-CoVs 和 δ-CoVs）的代表，并且可以检测到所有的内源性 CoV。在比较了这两种检测方法后，我们发现更新的检测法可靠地以高灵敏度检测了所有属的 CoV 病毒，而原始检测法的灵敏度较低。我们更新的 PCR 检测法是一种重要的工具，可以检测、监测和跟踪 CoV，以加强对宿主中病毒的监测。
