巢式聚合酶链反应技术在流感嗜血杆菌b型检测中的应用  

Identification of Haemophilus influenzae type b by nested polymerase chain reaction

作　　者：姜敏[1] 胡翼云[1] 高薇[1] 袁林[1] 沈叙庄[1] 俞桑洁[1] 杨永弘[1] 

机构地区：[1]首都医科大学附属北京儿童医院儿科研究所微生物室,北京100045

出　　处：《中国感染与化疗杂志》2006年第1期18-21,共4页Chinese Journal of Infection and Chemotherapy

基　　金：国家自然科学基金资助项目(30271377)

摘　　要：目的评价巢式聚合酶链反应(nested-PCR)技术在b型流感嗜血杆菌(Hib)检测中的应用价值,估计氨苄西林耐药Hi中b型的比率。方法对2000—2003年北京、上海、广州细菌监测项目中,呼吸道感染儿童鼻咽部分离培养的899株Hi,用E试验法检测氨苄西林耐药情况后,用nested-PCR与玻片凝集法对氨苄西林耐药菌株进行b型检测。结果检出74株氨苄西林耐药Hi。nested-PCR与玻片凝集法检测Hib的结果一致,74株氨苄西林耐药Hi中有2株为Hib,约占2.7%。结论nested-PCR b型荚膜检测方法可作为玻片凝集法的一个重要补充。3地4年内呼吸道感染的儿童鼻咽部携带的氨苄西林耐药Hi中,Hib的比率处于相对较低的水平。Objective The utility of eapsular typing by nested PCR method was evaluated. The prevalence of Haemophilus influenzae type b （Hib） strains among ampicillin-resistant isolates was determined. Methods A total of 899 H. infguenzae strains isolated from nasopharyngeal specimens of children with respiratory tract infection were investigated in Beijing, Shanghai and Guangzhou during the past four years from 2000 to 2003. Susceptibility to ampicillin was determined by E test. Nested PCR with primers specific to bexA gene and capsular type specific genes was established. Hib strains among resistant isolates were identified by nested-PCR based eapsular typing and slide agglutination serotyping （SATS） at the same time. Results Of the 74 H. influenzae strains resistant to ampicillin, 2 were positive by nested-PCR, and characterized as genotype b. The prevalence of Hib in ampicillin-resistant Hi isolates was about 2.7%. Conclusions Nested PCR based capsular typing as a supplement to slide agglutination serotyping, is useful for identifying Hib. The prevalence of type b strain among ampicillin-resistant H. influenzae isolated from nasopharyngeal specimens of children with respiratory tract infection in a defined geographic area during the past four vears is at relatively lover level.

关 键 词：B型流感嗜血杆菌 巢式聚合酶链反应 玻片凝集法 

分 类 号：R450[医药卫生—治疗学]