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Rapid Detection of Haemophilus influenzae and Haemophilus parainfluenzae in Nasopharyngeal Swabs by Multiplex PCR

查看全文 作　　者：TIAN Guo [1]Zhong;ZHANG Li [2]Juan;WANG Xiao [3]Lei;ZHANG [4]Li;LI Shu [5]Feng;GU Chang [5]Mei;SUN [5]Jian;CUI Bu [1]Yun 高影响力作者 机构地区：[1]State Key Laboratory for Infectious Diseases Control and Prevention, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China;[2]Department of Physiology and Pathophysiology, Basic Medical Sciences School of Capital Medical University, Beo'ing 100069, China;[3]The Third People's Hospital of Chengdu City, Chengdu 610031, Sichuan, China;[4]Chengdu Children Hospital, Chengdu 610017, Sichuan, China;[5]Feicheng Mining Group Company Health and Epidemic Prevention Station, Feicheng 2 71608, Shangdong, China高影响力机构 出　　处：《Biomedical and Environmental Sciences》索引2012年第25卷第3期,共5页高影响力期刊 摘　　要：Objective To establish multiplex PCR-based assays for detecting H.influenzae and H.parainfluenzae.And the PCR-based assays were applied to detect the carriage rates of H.influenzae and H.parainfluenzae in nasopharyngeal swab specimens which were collected from healthy children.Methods Multiplex primers for species-specific PCR were designed by using DNAstar soft based on the sequences of 16S rRNA genes from genus Haemophilus to detect H.influenzae and H.parainfluenzae.Results The sensitivity of the 16S rRNA PCR assay for detecting H.influenzae and H.parainfluenzae was 97.53% and 100% respectively,and the specificity was 95.89% and 96.63% respectively.Youden's Index on the ability to detect H.influenzae and H.parainfluenzae was 0.9342 and 0.9663 respectively.666 nasopharyngeal swab specimens were collected from healthy children.The detection rates of H.influenzae and H.parainfluenzae were 14.11% and 16.07% respectively by using isolation and culture methods.The detection rates of H.influenzae and H.parainfluenzae were 43.54% and 57.96% respectively by 16S rRNA PCR assays.The carriage rates of serotypes a,b,c,d,e,f and non-typeable isolates were 0% (0/666),0.15% (1/666),1.20% (8/666),0.15% (1/666),1.20% (8/666),1.80% (12/666),95.50% (636/666) respectively.Conclusion The multiplex PCR assays were very rapid,reliable and feasible methods for detection of H.influenzae and H.parainfluenzae in pharyngeal swab specimens which were compared to conventional isolation and culture methods.95.5% of H.influenzae strains in healthy children were nontypeable.The encapsulated or typable strains were mainly three serotypes which was c,e,and f serotype. 关 键 词：PCR快速检测 多重PCR检测 嗜血杆菌 副流感 RRNA基因序列 种属特异性 健康儿童 鼻
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