变性高效液相色谱分析肺炎支原体基因类型研究  

Genetype distribution of mycoplasma pneumoniae by denaturing high-performance liquid chromatography

作　　者：丁振尧 李红微[1] 郭美丽[1] 方全中 余宇龙 

机构地区：[1]浙江省苍南县人民医院儿科,325800

出　　处：《中国小儿急救医学》2014年第6期366-368,共3页Chinese Pediatric Emergency Medicine

基　　金：温州市科技计划项目（Y20130076）

摘　　要：目的 运用变性高效液相色谱（denaturing high-performance liquid chromatography,DHPLC）技术检测肺炎支原体（mycoplasma pneumoniae,MP）基因型,了解儿童MP流行状况.方法 对我院300例患儿鼻咽吸出物进行表型确证试验,采用PCR对标准菌株和MP表型阳性的临床菌株进行PCR扩增,扩增产物经DHPLC分析,通过与标准菌株色谱峰比对进行临床菌株基因分型.结果 300例患儿咽拭子经过24h培养后呈阳性110例.用特异引物扩增MP-129、MP-FH标准株和标本,分别得到2 280 bp和2580bp基因片段.110株临床菌株经DHPLC分析检测出107株是P1-Ⅰ型,均是1b亚型,3株是P1-Ⅱ型,均是2a亚型.结论 运用DHPLC技术检测出本院儿童发生MP感染以P1-Ⅰ、1b亚型为主.Objective To investigate the genetype distribution of mycoplasma pneumoniae（MP） by denaturing high-performance liquid chromatography（DHPLC）.Methods A total of 300 cases nasopharyngeal aspirate were collected from our hospital.The MP genes of standard strains and clinical specimens isolates were amplified by PCR followed by DHPLC and genetype determination.Results A total of 110 cases were positive after 24 hours fermentation from 300 cases with pharyngeal swab.By the specific primers of MP-129,MP-FH standard strain and specimens,2 280 bp and 2 580 bp gene fragments were made out respectively.One hundred and ten strains of clinical isolates were detected by DHPLC.One hundred and seven strains of P1-Ⅰ were 1b subtype,3 were type P1-Ⅱ which were all 2a subtype.Conclusion The genetype of MP infection in children from our hospital is P1-Ⅰ,1b subtype by using DHPLC technology.

关 键 词：变性高效液相色谱 肺炎支原体 基因分型 

分 类 号：R725.6[医药卫生—儿科]