重复序列PCR与多位点分型技术在白假丝酵母菌基因分型中的比较

发布时间：2018-01-30 04:42

本文关键词： 白假丝酵母菌多位点序列分型技术重复序列聚合酶链反应基因分型　出处：《上海交通大学学报(医学版)》2013年11期 　论文类型：期刊论文

【摘要】：目的应用多位点序列分型技术(MLST)和重复序列聚合酶链反应(REP-PCR)对临床分离出的白假丝酵母菌进行分型,并对两种方法作出应用评价。方法收集上海地区医院中真菌性阴道炎分离出的40株白假丝酵母菌,选择合适引物进行扩增,通过电泳比较分析获得REP-PCR分型。选择7对管家基因进行PCR反应并测序,将测序结果与标准序列进行比对,获得由7对管家基因组成的等位基因谱,最终得到相应的序列分型。结果 REP-PCR中Ca22-Ca22引物对得到电泳条带最优,40株白假丝酵母菌共分成7种REP-PCR型;MLST分型得出29种,且均为新型。结论 MLST分辨率较REP-PCR高,但由于REP-PCR分型方法更为快捷、经济,更适用于实验室大量菌株分型和临床分型。MLST则更客观,更适合用于进化学的研究及全球性流行病学的调查研究。
[Abstract]:Objective to identify the clinical isolates of Candida albicans by multilocus sequence typing technique (MLST) and repeat sequence polymerase chain reaction (REP-PCR). Methods 40 strains of Candida albicans isolated from hospitals in Shanghai were collected, and suitable primers were selected to amplify them. REP-PCR typing was obtained by electrophoretic analysis. Seven pairs of housekeeping genes were selected for PCR reaction and sequenced. The sequencing results were compared with standard sequences. The allelic spectrum composed of 7 pairs of housekeeping genes was obtained and the corresponding sequence typing was obtained. Results the Ca22-Ca22 primer pairs in REP-PCR obtained the best electrophoretic bands. 40 strains of Candida albicans were divided into 7 REP-PCR types. Conclusion the resolution of MLST is higher than that of REP-PCR, but the method of REP-PCR typing is faster and more economical. MLST is more suitable for laboratory and clinical typing. MLST is more suitable for chemical research and global epidemiological investigation.
【作者单位】：上海交通大学医学院附属瑞金医院检验科;上海交通大学医学院附属瑞金医院卢湾分院放射免疫检验科;
【基金】：上海市卫生局资助项目(2009239) 上海市科委生物医药资助项目(11411960500) 上海市医学重点专科建设项目(2012年A类21号)~~
【分类号】：R379
【正文快照】： 白假丝酵母菌是一种常见的条件致病菌。近年来,随着导管插管、免疫缺陷病和器官移植等问题逐渐增多,真菌感染及致死率都呈明显上升趋势。因此,针对白假丝酵母菌快速准确的分型对其致病性、耐药性和流行病学调查等均具有重要意义。目前,应用于真菌分型的分子生物学方法有很多,

【参考文献】