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Ｂｉｏ１１ｄＵＴＰ 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。
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.
