【作者】 陈敏； 廖万清； 吴绍熙； 姚志荣； 陈江汉； 徐红； 陈裕充；

【Author】 CHEN Min,LIAO Wan-qing,WU Shao-xi,YAO Zhi-rong,CHEN Jiang-han,XU Hong,CHEN Yu-chong Department of Dermatology,Changzheng Hospital,the Second Military Medical Universit

【机构】 第二军医大学长征医院皮肤科；

【摘要】 目的利用真菌核糖体基因内转录间隔区（ITS）序列分析和系统进化学分析来研究新生隐球菌上海变种的地位及变种间ITS序列的差异。方法采用真菌通用引物ITS1、ITS4,对12株新生隐球菌不同变种标准株的ITS片断进行PCR扩增和测序,结合GenBank等数据库中11株新生隐球菌标准株的ITS序列,用CLUSTAL W1.83软件多重比对分析序列的差别,MEGA3.1软件处理数据绘制系统进化树。结果新生隐球菌变种之间ITS序列存在明显差异,存在6种亚型;gattii变种ITS序列存在4种亚型;我国发现的新生隐球菌上海变种S8012与gattii变种在表型及分子生物学研究均存在显著的差异,但仍属于变种内差异,归入gattii变种的ITS C型;gattii变种RV20186归入gattii变种的ITS F型;新生隐球菌尿素酶阴性株D₂Y系一grubii变种突变菌株。结论我国发现的上海变种S8012归入gattii变种的ITS C型;原gattii变种RV20186归入gattii变种的ITS F型。更多还原

【Abstract】 Objective We extend our analysis of the ITS1-5.8S-ITS2 region sequence to all three varieties of Cryptococcus neoformans,including S8012 and 11 type strains,in order to evaluate the status of Cryptococcus neoformans var.S8012 and re-type Cryptococcus neoformans varieties.Methods Universal primers(ITS1 and ITS4) and PCR were used to amplify the DNA fragments of the ITS region and 5.8S rRNA gene,then it was analyzed by CLUSTAL W1.83 and MEGA3.1 software.Results We found that there exist six ITS types in the three varieties of Cryptococcus neoformans isolates and four ITS types(ITS type C,D,E and F) esist in Cryptococcus neoformans var.gattii.Cryptococcus neoformans var.S8012 belongs to Cryptococcus neoformans var.gattii ITS type C and Cryptococcus neoformans var.gattii strain RV20186 belongs to ITS type F.The urease negative strain D2Y was a mutant strain of grubii variety.Conclusion In addition to morphological differences, there are also distinctive differences based on the ITS typing.Cryptococcus neoformans var.shanghaiensis S8012 belongs to Cryptococcus neoformans var.gattii ITS type C.Six ITS types exist in the three varieties of Cryptococcus neoformans isolates.更多还原