SARS-CoV-2 Omicron变异株TaqMan探针法荧光定量RT-PCR的建立及应用
Chinese Journal of Microbiology and Immunology(2023)
Abstract
目的:开发和评估一种快速、简单和经济的替代测序的Omicron变异株荧光定量RT-PCR(RT-qPCR)。方法:针对SARS-CoV-2 ORF1ab保守区域、Omicron变异株S基因高频共同突变位点设计引物和TaqMan探针,建立Omicron株RT-qPCR检测方法,用经全基因组测序确定型别的样本进行验证,并对该方法的特异度和敏感度进行评估。结果:本研究建立的RT-qPCR分型法可以将Omicron变异株与包括早期A型流行株、Alpha和Delta变异株在内的SARS-CoV-2毒株进行区分,结果与全基因组测序结果一致,符合率为100.00%(28/28);与其他6种呼吸道病毒及柯萨奇病毒A组16型无交叉反应;该方法RNA标准品在10 9~10 3拷贝/μl之间呈良好的线性关系,相关系数 R2均大于0.99,检测敏感度为10 3拷贝/μl。 结论:本研究设计的针对Omicron变异株的RT-qPCR敏感度高且特异度好,在任何可以进行PCR检测的实验室中都容易开展,可极大地促进对Omicron变异株的传播监测。
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Key words
SARS-CoV-2,Omicron variants,RT-qPCR
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