多重PCR技术检测b型流感嗜血杆菌荚膜基因  

Detection of type b capsule gene of Haemophilus influenzae by multiplex PCR

作　　者：冯旭敏[1] 华春珍[1,2] 王超前[1] 项华冰[1] 来志超[1] 夏卫良[1] 王玲玲[1] 

机构地区：[1]杭州师范大学临床医学院,浙江杭州310036 [2]杭州师范大学附属医院

出　　处：《中国预防医学杂志》2012年第1期39-42,共4页Chinese Preventive Medicine

基　　金：浙江省科技计划项目(2006C33026)

摘　　要：目的应用多重PCR技术研究杭州地区儿童中分离的流感嗜血杆菌b型菌株的检出率。方法以玻片凝集法的血清分型为金标准,以流感嗜血杆菌(Hi)荚膜编码基因(bexA)和b型特异性荚膜基因序列设计引物,应用多重PCR技术对流感嗜血杆菌菌株进行荚膜基因检测。结果 2001-2002年和2006-2007年分离的399株Hi临床株中,血清分型显示不可分型株297株,占74.44%,可分型株102株,占25.56%。b型仅1株,构成比0.98%。多重PCR检测显示:102株玻片凝集法可分型的菌株中,101株bex A PCR结果阳性,1株阴性;297株不定型株中2株bex A阳性。敏感度99.02%;特异度99.33%。Hib检测结果显示b型1株,占有荚膜菌株0.99%,与血清分型结果一致。结论 bex A PCR联合针对b型特异性荚膜基因的多重PCR技术,对Hib检出率100%,克服了血清分型的弊端。Objective To detect Haemophilus influenzae type b strains isolated from children in Hangzhou by multiplex PCR methods. Methods All Haemophilus influenzae strains were analyzed by multiplex PCR. BexA-primers and b capsular type-specific gene primers were used as golden standard. Results Of all the 399 clinical strains isolated from children during the period from 2001-2002 and 2006-2007, 297 strains （74.44%） were nontypeable, while 102 strains （25.56%） were typeable and only 1 （0.25%） of them was type b, respectively. PCR results showed that of all the 102 strains which showed positive with the slide agglutination method, 101 were positive and only 1 was negative while 2 strains showed positive from the 297 strains which were nontypeahle. The results showed consistent with the serotype with the sensitivity of 99.02% and specificity of 99.33%. Conclusions With genotyping success rate and detection rate for Hib reaching 100%, the investigation overcame the shortcomings of serotyping by the technology of bex-PCR for b-type joint capsule gene specific multiplex PCR.

关 键 词：流感嗜血杆菌 b型 荚膜基因 儿童 多重PCR 

分 类 号：R440[医药卫生—诊断学]