不同核酸检测试剂盒检测环境样本禽流感病毒H5亚型的效果比较  

Comparison of the effectiveness of different nucleic acid detection kits in detecting avian influenza virus H5 subtypes in environmental samples

作　　者：林金思 杨淑欢 冯志锋 师舞阳[1] 吴衍恒[1] LIN Jinsi;YANG Shuhuan;FENG Zhifeng;SHI Wuyang;WU Yanheng(Department of Pathogenic Microorganisms and Biological Testing,Zhongshan Center for Disease Control and Prevention,Zhongshan,Guangdong 528400,China)

机构地区：[1]中山市疾病预防控制中心病原微生物与生物检验所,广东中山528400

出　　处：《热带医学杂志》2024年第2期181-184,共4页Journal of Tropical Medicine

摘　　要：目的 比较3种厂家检测禽流感病毒H5亚型的实时荧光定量逆转录聚合酶链反应(Real-time RT-PCR)试剂对环境标本的检测效果。方法 采用3个不同厂家的核酸检测试剂盒检测活禽市场环境样本中的H5亚型禽流感病毒,比较不同试剂检测结果检出率的差异,使用TA克隆技术,将扩增产物连接到克隆载体上并测序分析,获得反应产物的序列。结果 3种检测试剂中,试剂盒C检测H5的检出率比A高(18.3%vs. 8.4%),差异有统计学意义(χ^(2)=5.440,P=0.039)。对扩增产物片段序列进行分析发现,试剂A对标本核酸的H5扩增产物与KU042758.1序列的相似度最高,为100.00%,试剂C对标本核酸的H5扩增产物与CY091635.1序列的相似度最高,为97.92%,两者无明显的相似区域。结论 试剂C比试剂A对该批临床样本中H5的检出率更高,检出率的差异可能与不同试剂靶序列不一致以及试剂引物与样本核酸的匹配度有关。Objective To compare the detection efficiencies of real⁃time reverse transcription⁃polymerase chain reaction(Real⁃time RT⁃PCR)kits from three manufacturers for detecting avian influenza virus H5 subtype on environmental specimens.Methods Kits from three companies were used to detected H5 subtype avian influenza virus of the samples from live poultry market.Statistical methods were used to compare the difference in the detection rate of different kits.The PCR products were subjected to TA cloning and sequencing.Results Among the kits from three companies,the positive rate of H5 by using kits from company C was higher than that of company A(18.3%vs.8.4%),the difference was statistically significant(χ^(2)=5.440,P=0.039).TA cloning and sequencing showed that the sequence of PCR products of kit A shows highest similarity with sequence KU042758.1(100.00%).Sequence of PCR products of kit C shows highest similarity with sequence CY091635.1(97.92%).Conclusion Kit C had a higher detection rate of H5 in this batch of clinical samples than kit A,which might be caused by the inconsistency of the target sequence of different kits and the matching degree of the kit primer to the nucleic sample acid.

关 键 词：禽流感病毒 H5亚型 实时荧光定量逆转录聚合酶链反应 

分 类 号：R446.6[医药卫生—诊断学]