Updated and Validated Pan-Coronavirus PCR Assay to Detect All Coronavirus Genera.

Updated and Validated Pan-Coronavirus PCR Assay to Detect All Coronavirus Genera.
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DOI:
10.3390/v13040599
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发表时间:
2021-04-01
期刊:
Viruses
影响因子:
--
通讯作者:
van Doremalen N
van Doremalen N
中科院分区:
其他
文献类型:
--
作者:
Holbrook MG;Anthony SJ;Navarrete-Macias I;Bestebroer T;Munster VJ;van Doremalen N

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来自野生动物宿主的冠状病毒(CoV)溢出事件可导致轻度至重度人类呼吸道疾病。这些溢出事件表明,必须检测在宿主物种中传播的已知和新型冠状病毒,并确定冠状病毒的流行和分布,从而改进溢出事件的预测,或密切监测人-库界面的位置。为了提高检测所有流行属和毒株的可能性,我们修改了Watanabe等人在2010年发表的引物,以生成半巢式泛冠状病毒PCR检测。对4种冠状病毒属(α- cov、β- cov、γ- cov和δ- cov)的代表进行了检测,并对所有内部冠状病毒进行了检测。在比较两种检测方法后,我们发现更新后的检测方法可靠地检测到所有属的冠状病毒,灵敏度高,而原始检测方法的灵敏度较低。该方法是检测、监测和跟踪冠状病毒的重要工具,可加强对宿主病毒的监测。
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.
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