多位点序列分型技术在热带念珠菌中的应用研究

中国真菌学杂志　 2018, Vol. 13 　Issue (6): 321-326.

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多位点序列分型技术在热带念珠菌中的应用研究

王影¹, 项明洁², 刘锦燕³, 叶书来¹, 周馨¹

1. 中国科技大学附属第一医院检验科, 合肥 230001;
2. 上海交通大学附属瑞金医院检验科, 上海 200025;
3. 上海交通大学附属瑞金医院卢湾分院放免检验科, 上海 200020

收稿日期:2018-04-25 出版日期:2018-12-28 发布日期:2018-12-28
通讯作者: 周馨,E-mail:zhouxin1806@126.com E-mail:zhouxin1806@126.com
作者简介:王影,女(汉族),硕士,初级检验师.E-mail:sjdwangying@163.com

The study of multilocus sequence typing in Candida tropicalis

WANG Ying¹, XIANG Ming-jie², LIU Jin-yan³, YE Shu-lai¹, ZHOU xin¹

1. Department of Clinical Laboratory, The First Affiliated Hospital of University of Science and Technology of China, Hefei 230001, China;
2. Department of Laboratory Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China;
3. Department of Laboratory Medicine, Ruijin Hospital Luwan Branch, Shanghai Jiao Tong University School of Medicine, Shanghai 200020, China

Received:2018-04-25 Online:2018-12-28 Published:2018-12-28

摘要/Abstract

摘要：

目的探讨上海瑞金医院临床分离的热带念珠菌MLST（Multilocus sequence types，MLST）型别。方法连续收集上海市瑞金医院临床分离的热带念珠菌菌株，用PCR技术扩增热带念珠菌的6对管家基因，并对扩增片段进行纯化测序后与热带念珠菌MLST数据库比对得到临床菌株的二倍体序列型别DSTs（Diploid sequence types，DSTs），再利用eBURST V3软件对结果进行分析，用MEGA 6.0软件建立系统发育树，最后对部分临床菌株进行药物敏感性实验，探讨菌株耐药与基因型别之间的关系。结果临床分离的92株热带念珠菌分属于47个DSTs，其中15个DSTs是已经报道的，32个DSTs在本次研究中首次得到；DST507和DST376是本次研究中的优势型别，且属于同一个克隆簇。药敏实验显示属于优势型别的27株临床菌株中21株为唑类药物耐药菌株。结论 MLST技术是一种高效的基因分型技术；DST507和DST376是上海瑞金医院热带念珠菌的优势型别；唑类药物耐药菌株表现为DST507和DST376的聚集分布。

关键词: 热带念珠菌, 基因分型, MLST, DSTs

Abstract:

Objective To study the dominant genotypes of multilocus sequence typing (MLST) about Candida tropicalis in Shanghai, China. Methods The clinical Candida tropicalis isolates were collected from Ruijin Hospital, Shanghai, China, and identified using morphological analysis on CHROMagar Candida and carbohydrate assimilation tests by API 20C AUX. Six housekeeping genes of Candida tropicalis were amplified and sequenced, and then the allelic profiles of the six gene sequences were obtained from the Candida tropicalis MLST database. The results were further analysed using the eBURST V3 package to determine the MLST clonal clusters. The phylogenetic tree was established using MEGA 6.0 software. Then the drug susceptibility tests were conducted to explore the relationship between strain resistance and genotype. Results There were 47 DSTs were obtained from 92 clinical Candida tropicalis isolates, from which 32 DSTs were new to the MLST database. DST507 and DST376 were the dominant genotypes, and they belong to the same clonal cluster. 21 of 27 clinical strains belonging to the dominant genotypes were azole-resistant strains. Conclusions MLST was a high-efficiency genotyping method for Candida tropicalis. DST507 and DST376 were the dominant genotypes to Candida tropicalis. It is necessary to monitor the spread of azole-resistant Candida tropicalis, especially clonal dissemination of DST507 and DST376 in the future.

Key words: Candida tropicalis, genotyping, MLST, DSTs

中图分类号:

R379.4

王影, 项明洁, 刘锦燕, 叶书来, 周馨. 多位点序列分型技术在热带念珠菌中的应用研究[J]. 中国真菌学杂志, 2018, 13(6): 321-326.

WANG Ying, XIANG Ming-jie, LIU Jin-yan, YE Shu-lai, ZHOU xin. The study of multilocus sequence typing in Candida tropicalis[J]. Chinese Journal of Mycology, 2018, 13(6): 321-326.

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多位点序列分型技术在热带念珠菌中的应用研究

The study of multilocus sequence typing in Candida tropicalis

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