液相芯片技术在新冠病毒和流感病毒检测中的应用  

Detecting of SARS-CoV-2 and influenza virus by using liquichip technology

作　　者：周有良 胡春凌 徐俊 陶莉 陈凯 孙玉莲 葛斌鹏 ZHOU Youliang;HU Chunling;XU Jun;TAO Li;CHEN Kai;SUN Yulian;GE Binpeng(Suzhou Customs,Suzhou,Jiangsu 215011,China;不详)

机构地区：[1]苏州海关,江苏苏州215011 [2]深圳市普捷思生物科技有限公司 [3]苏州市姑苏区民政和卫生健康局

出　　处：《中国国境卫生检疫杂志》2023年第5期405-410,共6页Chinese Journal of Frontier Health and Quarantine

基　　金：南京海关科研项目(2022KJ14)。

摘　　要：目的 建立一种液相芯片检测方法,用于检测新冠病毒和流感病毒。方法 从NCBI数据库分别下载新冠病毒、甲型流感病毒、乙型流感病毒全基因组序列,在保守区域分别设计扩增引物和检测探针,以各病毒标准品质粒为模板,建立多重PCR扩增体系;将核酸探针偶联至荧光编码微球,建立液相芯片检测体系,并对方法进行评价。结果 基于多重PCR和液相芯片技术建立的新冠病毒和流感病毒的检测方法能够同时检测新冠病毒N基因、ORF1ab基因和S基因,甲型流感病毒M基因和乙型流感病毒NS基因及人类看家基因ACTB基因。检测新冠病毒N基因、ORF1ab基因、S基因的灵敏度为1×10^(4)拷贝/反应,甲型流感病毒M基因、乙型流感病毒NS基因的灵敏度为1×10^(3)拷贝/反应;可准确检测新冠病毒N基因、ORF1ab基因和S基因,甲型流感病毒M基因和乙型流感病毒NS基因及人类看家基因ACTB基因片段,没有出现相互干扰的现象。结论 建立的方法能同时检测新冠病毒、甲型流感病毒和乙型流感病毒,可作为新冠病毒及流感病毒感染的诊断及流行病学调查的方法。Objective To establish a liquichip method which can be used for detecting severe acute respiratory syndrome coronavirus 2(SARS-CoV-2) and influenza virus.Methods The genomic sequence of SARS-CoV-2,influenza A virus and influenza B virus were download from NCBI database.The amplification primers and detection probes were designed in the conserved regions of each virus.Multiplex PCR amplification system was established by using the standard plasmids of each virus as the template.The nucleic acid probes were coupled to fluorescent microspheres,and liquichip detection system was established and evaluated.Results The established method based on multiplex PCR and liquichip technology could detect N gene,ORF1ab gene and S gene of SARS-CoV-2,and M gene of influenza A virus,NS gene of influenza B virus,and human housekeeping gene ACTB gene simultaneously.The deterction sensitivity of N gene,ORF1ab gene and S gene of SARS-CoV-2 was 1×10^(4)copies/reaction,the detection sensitivity of M gene of influenza A virus and NS gene of influenza B virus was 1 ×10^(3)copies/reaction.It could accurately detect N gene,ORF1ab gene and S gene of SARS-CoV-2,M gene of influenza A virus and the NS gene of influenza B virus,and human housekeeping gene ACTB gene fragments without any interference.Conclusion This method could be used for detecting SARS-CoV-2,influenza A virus and influenza B virus simultaneously,and could be used as a method for diagnosis and epidemiological investigation of SARS-CoV-2 and influenza virus infection.

关 键 词：新冠病毒 甲型流感病毒 乙型流感病毒 液相芯片 多重PCR 

分 类 号：R183.5[医药卫生—流行病学]