多态性微卫星标记技术与重复序列PCR在近平滑念珠菌基因分型中的比较  

Comparison on genotyping of Candida parapsilosis by polymorphic microsatellite marker technology and repetitive sequence-based PCR

作　　者：李贞[1] 董丹凤[1] 章黎华[1] 江岑[1] 彭奕冰[1] 

机构地区：[1]上海交通大学医学院附属瑞金医院检验科,上海200025

出　　处：《检验医学》2015年第10期1017-1020,共4页Laboratory Medicine

基　　金：国家自然基金面上项目(81371873);国家自然基金资助项目(81301462)

摘　　要：目的比较多态性微卫星标记（PMM）技术与重复序列聚合酶链反应（PCR）在近平滑念珠菌基因分型中的应用。方法收集2012年3月至2013年11月间上海5家医院临床分离的近平滑念珠菌55株,经内转录间隔区序列分析（ITS）进一步鉴定菌种,共有狭义近平滑念珠菌43株,之后采用重复序列PCR和PMM技术进行基因分型,比较2种分型方法的结果和效率。结果 43株狭义近平滑念珠菌经重复序列PCR分型只有1种基因型;经PMM技术分型有22种基因型,以Ⅰ型（10株）、Ⅱ型（6株）为主,Ⅲ~Ⅶ型见于2或3株菌,其余15种基因型均只有1株菌。结论 PMM技术用于狭义近平滑念珠菌的基因分型,分辨率高、重复性好,适用于狭义近平滑念珠菌基因组微进化的监测,而重复序列PCR不适用于狭义近平滑念珠菌的基因分型。Objective To evaluate the application of polymorphic microsatellite marker（ PMM） technology and repetitive sequence-based polymerase chain reaction（ PCR） for genotyping of Candida parapsilosis. Methods From March 2012 to November 2013,55 Candida parapsilosis isolates were collected from 5 hospitals in Shanghai,which were further identified as Candida parapsilosis sensu stricto（ 43 isolates） by amplifying and sequencing with internal transcribed sequence（ ITS）. All the isolates were genotyped by repetitive sequence-based PCR and PMM technology.Finally,the results and efficacy of these 2 genotyping methods were compared. Results The 43 Candida parapsilosis sensu stricto isolates had only 1 genotype by repetitive sequence-based PCR. PMM technology generated 22 genotypes.The major genotypes were Type Ⅰ（ 10 isolates） and Type Ⅱ（ 6 isolates）,whereas Type Ⅲ-Ⅶ were observed in 2 or3 isolates. Another 15 genotypes were observed in only 1 isolate. Conclusions PMM technology is an effective genotyping method for Candida parapsilosis sensu stricto with high resolution and repeatability,which could be used to determine the micro-evolutionary variations. However,repetitive sequence-based PCR is not suitable for the genotyping of Candida parapsilosis sensu stricto.

关 键 词：近平滑念珠菌 基因分型 多态性微卫星标记技术 重复序列聚合酶链反应 

分 类 号：R379.4[医药卫生—病原生物学]