白假丝酵母荧光定量PCR鉴定方法的建立  

Establish a real-time PCR method to rapid identify Candida albicans

作　　者：许凡凡 贺丹[2,3] 孙晓红[2,3,4] 王丽[2,3] 高嵩[2,3] 盛辉[1] 

机构地区：[1]吉林大学第一医院呼吸科,吉林长春130021 [2]吉林大学基础医学院病原生物学系 [3]吉林大学真菌研究中心.教育部人兽共患病重点实验室,吉林长春130021 [4]北华大学基础医学院,吉林吉林132013

出　　处：《中国实验诊断学》2015年第4期528-531,共4页Chinese Journal of Laboratory Diagnosis

基　　金：吉林省科技发展计划项目(20110453;20130522014JH)

摘　　要：目的真菌感染发病率和死亡率逐年增高,亟需建立主要病原真菌快速、敏感、特异的鉴定方法,对感染的早期诊治至关重要。方法以真菌线粒体COⅡ基因为靶标,设计白假丝酵母的特异性引物,建立荧光定量PCR鉴定方法,分析其灵敏度、特异性及重复性。结果该方法检测的灵敏度可达1×102copies/μl(相当于10-7 ng/μl),熔解曲线显示具有单一峰,且仅有白假丝酵母出现扩增曲线,其他种、属真菌均未见扩增。且具有较好的重复性和稳定性。结论建立的白假丝酵母荧光定量PCR鉴定方法灵敏度高、特异性强、重复性和稳定性好,适于临床真菌感染微量标本的快速检测。该研究为真菌核酸快速检测试剂盒的开发奠定了基础。Objective The morbidity and mortality of fungal infections increased these years.It is necessary to es-tablish a rapid,sensitive and specific identification method which is essential for early diagnosis and treatment of the in-fections.Methods Specific primers of Candida albicans were designed according to the sequences of fungal mitochon-drial COⅡ gene.And a real-time PCR method was established to identify Candida albicans .The sensitivity,specificity and repeatability of this identification method were analyzed.Results Sensitivity of the method was up to 1 × 102 copies/μl （equivalent 10-7 ng/μl）.There was a single peak of dissociation curve.Only Candida albicans was ampli-fied,other genus and speciesof fungi were not amplified.The method had good repeatability and stability.Conclusion The established real-time PCR method of Candida albicans has higher sensitivity and specificity,good repeatability and stability,which is suitable for rapid diagnosis with trace specimens of fungal infections in clinic.This study will lay the foundation for the development of rapid diagnostic kit of fungi.

关 键 词：白假丝酵母 荧光定量PCR 鉴定 COⅡ基因 

分 类 号：R440[医药卫生—诊断学]