张军民 周贵民 谢玲 杨瑞馥 张军民 周贵民 谢玲 100853 北京，解放军总医院微生物科；杨瑞馥 军事医学科学院微生物流行病学研究所 中华医学检验杂志 1999 0 22 6


    关键词：聚合酶链反应 杂交 念珠菌属 期刊 zhjyyxzz 0 论著摘要 fur -->


    

【摘要】 目的 研究以非培养为基础的快速检测与鉴定念珠菌的方法。方法 将白色念珠菌、热带念珠菌、光滑念珠菌种特异探针加尾后固定在硝酸纤维素膜上；用氯化苄法提取念珠菌DNA，采用真菌特异通用引物PCR扩增DNA，在扩增过程中用Bio-11-dUTP标记扩增产物，然后分别与白色念珠菌、热带念珠菌、光滑念珠菌特异探针杂交。结果 对14种念珠菌扩增均为阳性，对3种常见细菌扩增均为阴性。3种念珠菌只与其对应探针杂交。结论 该方法简便、快速、准确，适于临床实验室快速检测与鉴定念珠菌。

Rapid detection and identification of Candida species by PCR-reversed dot hybridization

    ZHANG Junmin^* , ZHOU Guimin, XIE Ling, et al.

    ^* Department of Microbiology, Chinese PLA General Hospital, Beijing 100853

【 Abstract 】 Objective To establish a non-culture method for detection and identification of Candida isolates from clinical specimens.Methods The species-specific probes to identify three Candida species, C.albicans,C.tropicalis and C.glabrata, were added homopolymer tails and then inoculated on a nitrocellulose filter. The benzyl chloride method was used for extracting template DNA from Candida isolates. The DNA from each isolate was amplified by PCR with the fungus-specific universal primers, and the PCR products labeled with bio-11-dUTP during amplification were hybridized with the species-specific probe.Results All 14 Candida species were amplified by PCR, and non-specific amplificatin for 3 bacteria species was found. The probes reacted with C.albican, C.tropicalis and C.glabrata didn′ t react with other Candida labeled DAN.Conclusion PCR-reversed dot hybridization is rapid and simple for detection and identification of medically important Candida species.

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