MLMT与MLST在新生隐球菌基因分型中的应用研究  

Application of genotypes in Cryptococcus neoformance strains based on MLMT and MLST

作　　者：朱键 曹佳非 康颖倩 三上襄 Wieland Meyer ZHU Jian;CAO Jiafei;KANG Yingqian;SAN Shangxiang;Wieland Meyer(Department of Gastroenterology,the Second People's Hospital of Guiyang&the Affiliated Jinyang Hospital of Guizhou Medical University,Guiyang 550081,Guizhou,China;Department of Gastroenterology,the Affiliated Hospital of Guizhou Medical University,Guiyang 550004,Guizhou,China;Graduate School,Guizhou Medical University,Guiyang 550004,Guizhou,China;Medical Mycology Research Center,Chiba University,Chiba 260-8673,Japan;Westmead Institute for Medical Research,the University of Sydney,NSW2145,Sydney,Australia)

机构地区：[1]贵阳市第二人民医院&贵州医科大学附属金阳医院消化内科,贵州贵阳550081 [2]贵州医科大学附属医院消化内科,贵州贵阳550004 [3]贵州医科大学研究生院,贵州贵阳550004 [4]千葉大学医学真菌研究中心,日本千葉县260-8673 [5]悉尼大学Westmead医学研究所,澳大利亚悉尼NSW2145

出　　处：《贵州医科大学学报》2024年第1期84-89,共6页Journal of Guizhou Medical University

基　　金：贵州省卫生健康委员会科学技术基金项目(gzwkj2013-1-055)。

摘　　要：目的 探讨多位点微卫星灶分型(MLMT)和多位点序列分型(MLST)在新生隐球菌(C.neoformans)基因分型中的应用。方法 提取22株来源于国内及日本已明确为新生隐球菌的菌株DNA,经MLMT及MLST相应位点的引物进行扩增、测序;计算各微卫星灶(CNG1、CNG2及CNG3)序列中简单重复序列的重复数,确定菌株MLMT的基因型;将本研究菌株的MLST中各位点序列和网上数据库相应序列进行对比,获取该菌株的MLST基因型;比较两种方法在分型中分辨能力。结果 在22株菌中,有16株的基因型为MLMT17/MLST5(16/22,72.73%),MLMT将所有菌株分为7型,而MLST分为4型;根据MLST分型,MLMT17、16、34及39的菌株为MLST5;MLMT2、14、29相对应MLST66、32及31;两种方法的分型分辨能力比较,差异无统计学意义(P=0.488)。结论 MLMT-17/MLST 5是我国和日本的新生隐球菌常见基因型,包含CNG1、CNG2及CNG3位点的MLMT可作为MLST的替代或补充。Objective To investigate the application of Multilocus Microsatellite Typing(MLMT)and Multilocus Sequence Typing(MLST)in the genotype of Cryptococcus neoformans(C.neoformans).Methods The DNA of 22 C.neoformans strains from China and Japan was extracted,amplified and sequenced by MLMT and MLST primers.The number of short tandem repeats in each microsatellite locus(CNG1,CNG2,and CNG3)sequence was calculated to determine the genotype of MLMT.The genotype of the strain was obtained by comparing the strain locus sequence of MLST in the study with the corresponding sequence of the online database.Genotyping resolution of the two methods was compared by SPSS 19.0 statistical software.Results Among all 22 strains,16 were identified as MLMT17/MLST5(16/22,72.73%).All strains were determined as 7 types by MLMT and 4 types by MLST.According to MLST,MLMT 17,16,34,and 39 were classified into MLST5;MLMT2,14,and 29 were corresponding to MLST 66,32,and 31.There was no significant difference of genotyping ability between the two methods(P=0.488).Conclusion MLMT-17/MLST 5 is a common genotype of C.neoformans in China and Japan.MLMT containing CNG1,CNG2,and CNG3 microsatellite locus can be used as a substitute or supplement method of MLST for C.neoformans.

关 键 词：新生隐球菌 多位点微卫星灶分型 多位点序列分型 

分 类 号：R446.5[医药卫生—诊断学]