随着移植肿瘤和免疫缺陷患者的增多侵袭性真菌感染invasive.docVIP

PCR-高分辨率溶解曲线（HRM）快速鉴定常见念珠菌 郭宁辉，刘 静,丁 雪，任 微，王 璐，褚美玲，孟冬娅，薛文成 摘要：目的 探讨建立一种快速鉴定念珠菌的PCR-高分辨率溶解曲线（High Resolution Melting，PCR-HRM）分析方法，为临床抗真菌感染治疗的及时、合理用药提供帮助。方法 共收集117株临床分离的9种念珠菌以念珠菌ITS2区DNA序列为靶基因进行PCR-HRM分析，以测序结果为金标准评估PCR-HRM分析方法鉴定念珠菌菌种的可靠性。结果 临床分离的常见菌种如白色念珠菌、热带念珠菌、光滑念珠菌、近平滑念珠菌、克柔念珠菌、葡萄牙念珠菌等菌种，均具有独特的PCR-HRM特征性曲线。结论 PCR-HRM分析方法有可能成为临床分离念珠菌的快速鉴定方法，但在检测结果的自动判定等方面还需要进一步开发研究。 关键词：高分辨率溶解曲线（HRM）；酵母样真菌；快速鉴定； 中图分类号：R446 文献标志码：A 基金项目：辽宁省科技攻关计划（No.2011225021） 作者单位：121001锦州市 辽宁医学院沈阳军区总医院研究生培养基地（郭宁辉）；沈阳军区总医院检验科（刘 静，褚美玲,丁 雪，任 微，王 璐，孟冬娅，薛文成）（郭宁辉与刘静同等贡献） 通讯作者：薛文成，电子信箱189.cn Rapid identification of Candida isolates by use of PCR and High-Resolution Melt Analysis Guo Ninghui*, Liu Jing, Chu Meiling, Ding Xue, Ren, Wei Wang Lu, Meng Dongya, XUE Wencheng （*Postgraduate Training Base of General Hospital of Shenyang Military Command, Liaoning Medcial University,Jinzhou 121001,China; Clinical Labratory,General Hospital of Shenyang Military Command,Shenyang 110016,China） Corresponding author: Xue Wencheng, Email189.cn 【Abstract】 Objective To develop a new diagnostic method based on real-time polymerase-chain reaction high-resolution melting analysis (PCR-HRM) that detects yeasts, to assist in an timely antifungal therapy and appropriate drug treatments. Methods All 9 Candida includes 117 isolates analysed in this study were obtained from the clinical, and the analysis of the PCR-HRM is target at the internal transcribed spacer 2 (ITS2) DNA sequencing of the yeast, the gene sequencing as a “golden standard” to evaluate the reliability of the PCR-HRM proposal to identify the yeast at the level species. Reasults The frequently isolates from the clinical comprises C. albicans, C. tropicalis, C. glabrata, C. parapsilosis, C.krusei, C.lusitaniae ect,and they all have a unique PCR-HRM characterisitic curve. Conlusions The PCR-HRM proposal as a diagnostic approach will be able to detect the yeast isolates from the clinical, but the future research is needed at