共有序列简并杂合寡核苷酸引物聚合酶链反应在呼吸道副黏病毒科病毒诊断中的应用  

Consensus-degenerate hybrid oligonucleotide primer polymerase chain reaction ——A powerful technique for the identification of Paramyxoviridae in clinical specimens

作　　者：赵百慧[1] 王春[2] 沈佳仁[1] 俞雪莲[1] 高烨[1] 滕峥[1] 朱兆奎[1] 储维[3] 宋黎黎[3] 张泓[2] 张曦[1] 

机构地区：[1]上海市疾病预防控制中心,上海200336 [2]上海交通大学附属上海市儿童医院,上海200040 [3]上海市黄浦区疾病预防控制中心,上海200023

出　　处：《微生物与感染》2013年第2期89-95,共7页Journal of Microbes and Infections

基　　金：上海市卫生局青年科研项目(20124Y184);上海市公共卫生人才培养计划(GWDTR201201);上海市公共卫生重点学科建设(12GWZX0801)

摘　　要：采用共有序列简并杂合寡核苷酸引物聚合酶链反应(CODEHOP PCR)体系和商品化RV12试剂盒对急性呼吸道感染患儿下呼吸道标本中的副黏病毒科病毒进行检测,比较CODEHOP PCR与RV12试剂盒检测结果的符合率和敏感度,观察CODEHOP PCR在临床呼吸道标本中对副黏病毒诊断的应用价值,进一步探讨具备检测已知呼吸道病毒和未知新病毒特点的CODEHOP PCR在呼吸道疾病谱和未知潜在呼吸道病毒诊断中的推广价值。采集2011年上海市儿童医院因急性呼吸道感染住院的患儿下呼吸道标本572份,分别采用2种方法对副黏病毒进行检测。CODEHOP PCR检测出阳性标本113例,阳性率为19.76%;RV12试剂盒检测出阳性标本102例,阳性率17.83%;2种方法检测符合率为85.39%。以10倍倍比稀释呼吸道合胞病毒A(RSVA)感染的细胞收获液,检测结果显示,CODEHOP PCR和RV12试剂盒检测下限分别为10-8和10-6。CODEHOP PCR具备简便、易操作和测序精度高等特点,适合疾病预防系统和临床检验科使用。该方法与一些新兴的高通量快速分子诊断技术结合,可提高检测灵敏度,具有高通量、快速检测和筛查未知新病毒的优势,在呼吸道病原体检测领域值得进一步优化和推广。The present paper aims to compare the sensitivity, specificity and accordance rate of consensus- degenerate hybrid oligonucleotide primer polymerase chain reaction （CODEHOP PCR） technique and commercial kit in diagnosis of paramyxovirus infection in clinical specimens and to further evaluate the value of CODEHOP PCR assay in detection of known and novel respiratory viruses. A total of 572 specimens from lower respiratory tract were collected from Children＇ Hospital of Shanghai inpatients with acute respiratoryinfections during 2011. These specimens were analyzed by CODEHOP PCR and commercial RV12 kit for the detection of paramyxoviruses, including known parainfluenza virus type 1 （PIV-1）, PIV-2, PIV-3, respiratory syncytial virus A （RSVA）, RSVB, human metapneumovirus （HMPV） and novel viruses. The results showed that 102 out of 572 specimens （19.76%） were detected positive by the RV12 kit, and 113 were detected positive by CODEHOP PCR. The accordance rate of the two methods was 85.39% . CODEHOP PCR is more sensitive than RV12 kit in detecting RSVA infection in cell culture. The study suggests that CODEHOP PCR is a powerful technique for the identification of paramyxoviruses in clinical specimens. It can be optimized by combining with some new techniques, and is worthy of application in the future.

关 键 词：共有序列简并杂合寡核苷酸引物聚合酶链反应 副黏病毒科 新呼吸道病毒 

分 类 号：R440[医药卫生—诊断学]