浙江省儿童鼻咽吸出物标本肺炎支原体基因分型研究  

Genotyping of Mycoplasma pneumoniae in nasopharyngeal aspirate from children with pneumonia in Zhejiang Province

作　　者：张立[1] 陈志敏[1] 沈征[2] 李淑娴[1] 徐丹[1] 唐兰芳[1] 

机构地区：[1]浙江大学医学院附属儿童医院呼吸科,杭州310003 [2]浙江大学医学院附属儿童医院中心实验室,杭州310003

出　　处：《中华儿科杂志》2011年第10期750-754,共5页Chinese Journal of Pediatrics

基　　金：基金项目：国家自然科学基金（81070004,30872798）;浙江省教育厅重点科研计划项目（20070248）;浙江省教育厅课题（Y200804505）

摘　　要：目的对浙江省肺炎支原体感染儿童鼻咽吸出物标本进行肺炎支原体基因分型研究，旨在了解肺炎支原体亚型的流行状况及其与临床的关系。方法采用基于肺炎支原体P1基因的聚合酶链反应（PCR）和限制性片段长度多态性分析技术，对2009年2—12月浙江大学医学院附属儿童医院呼吸科因肺炎支原体肺炎住院患儿的鼻咽吸出物标本300例进行肺炎支原体基因亚型检测并选取部分P1-I型和P1-Ⅱ型标本进行测序，比较不同亚型肺炎支原体感染患儿的临床资料。结果297例肺炎支原体感染临床标本显示肺炎支原体P1-I型特征性条带，3例显示P1-Ⅱ型特征性条带。所有P1-I型临床标本均显示Ib亚型条带，3例P1-Ⅱ型均显示Ⅱa亚型条带。测序结果显示P1-I型肺炎支原体P1基因第208位点出现了G—A的同义点突变。3例P1-Ⅱ型肺炎支原体肺炎患儿均合并肝功能损害，平均住院时间和发热时间比P1-I型长，但未达统计学差异。结论临床标本可直接用于肺炎支原体基因分型。研究期间浙江省发生的肺炎支原体感染以P1-I型、Ib亚型为主。肺炎支原体亚型与病情的关系有待进一步研究。Objective The P1 protein of Mycoplasma pneumoniae （MP） plays an important role in the pathogenesis of MP pneumonia. It mediates the attachment of the pathogen to host cells and elicits a strong humoral immune response during infection. In early studies, only two types of MP P1 genes were assumed to exist. Later, eight subtypes of MP P1 genes and some variations of P1 gene were reported. However, there are no related reports in China until now. This study aimed to understand epidemiology of MP subtype in Zhejiang province, China, as well as the relationship between MP subtype and clinical severity of MP pneumonia. Method Clinical samples were collected by nasopharyngeal aspiration from children with MP pneumonia hospitalized in the Children＇s Hospital of Zhejiang University School of Medicine from February to December in 2009. P1 gene fragment was amplified by using PCR method （with primers of ADH1/ADH2 and ADH3/ADH4, respectively）. Then ADH1/ADH2-generated fragments were digested with I-Iae Ⅲ, Hpa Ⅱ , Sau3A, and the ADH3/ADH4-generated fragments digested with Hae Ⅲ, Sau3A, Hha I , Rsa I. The MP P1 subtypes were determined based on resulting fragments. Part of samples were selected for sequencing. The clinical data of different MP subtype pneumonia were compared. Result A total of 300 hospitalized children with MP pneumonia were enrolled in this study. All the samples produced specific bands for MP P1 gene after PCR with primers of ADH1/ADH2 and ADI,H3/ADH4 respectively. By restrictive fragment length polymorphism analysis, 297 clinical specimens showed the characteristic band patterns for P1 type 1 identical to Mp129, and only 3 clinical specimens showed the characteristic band pattern for P1 type 2 identical to MP-FH. All P1 type 1 and P1 type 2 showed the samesubtype bands respectively, as subtype lb and 2a. After sequencing, one synonymous point mutation in P1 type 1 was identified relative to the MP129 P1 sequence at nucleotide position （nt） 208 （G→A）. Three cases with P1 type 2 MP p

关 键 词：支原体 肺炎 基因型 多态性 限制性片段长度 儿童 

分 类 号：R192.3[医药卫生—卫生事业管理]