不同RT-PCR扩增试剂盒检测甲肝病毒效果评价  

Effective Evaluation of Different RT-PCR Amplification Kits for Detection of Hepatitis A Virus

作　　者：齐欣 曲世超 田卓 麻丽丹 崔妍 徐文英 王煜 QI Xin;QU Shi-Chao;TIAN Zhuo;MA Li-Dan;CUI Yan;XU Wen-Ying;WANG Yu(Technical Center of Dalian Customs,Dalian 116001;Dandong Customs,Dandong 118001)

机构地区：[1]大连海关技术中心,大连116001 [2]丹东海关,丹东118001

出　　处：《中国口岸科学技术》2020年第7期43-52,共10页China Port Science and Technology

基　　金：辽宁省自然基金项目(2019-MS-395)资助。

摘　　要：讨论评价ABI公司、天根公司和Roche公司扩增试剂盒用于检测食品中甲肝病毒结果的差异,为甲肝病毒扩增检测提供便捷、准确的技术参考。在食品样品中添加盔甲病毒作为病毒富集的过程质控,甲肝减毒疫苗作为阳性对照。根据一步法RT-PCR检测人工污染样品的Ct值对三种扩增试剂盒的扩增效率、特异性和灵敏度进行评价和数据分析。ABI公司扩增试剂盒扩增效率最高(EFF%:91.584),灵敏度最高(0.314CCID50/μL),三种扩增试剂盒均具有良好的特异性。采用盔甲病毒作为过程控制病毒进行食品中甲肝病毒的检测,为甲肝病毒的质量控制提供分析依据。ABI公司扩增试剂盒在总体性能上优于其他两个厂家,更适用于各级实验室甲肝病毒的日常检测工作,为一线检测工作提供最佳的扩增方案及质量控制措施。The aim of this work is to discuss and evaluate the detection difference of Hepatitis A virus from food by three various kinds of amplification kits produced by ABI,Tiangen and Roche,to provide a convenient and accurate technical reference for Hepatitis A virus amplification test.Armored virus were added into the food samples as the process quality control of virus enrichment and hepatitis A attenuated vaccine with different titers was used as a positive control.The cycle threshold(Ct)of artificial contaminated negative samples was detected by one-step RT-PCR and used as the index for evaluation and analysis of extraction efficiency,detection specificity and sensitivity of three different amplification kits.Although all the three amplification kits have good detection specificity,the ABI amplification kit shows highest extraction efficiency(EFF%:91.584)and detection sensitivity(0.314 CCID50/μL).Armored virus was used as a process control virus to detect hepatitis a virus in food,providing an analytical basis for quality control of hepatitis A virus.The performance of ABI amplification kit is superior to the other two kits.It is more suitable for daily detection of hepatitis a virus in lab,providing an excellent amplification solution and quality control measurement for first-line testing.

关 键 词：扩增试剂盒 甲肝病毒 盔甲病毒 评价 

分 类 号：R512.61[医药卫生—内科学]