微孔板双探针杂交法快速鉴别都柏林念珠菌和白念珠菌

目的 建立微孔板双探针杂交法，快速鉴别都柏林念珠菌和白念珠菌，并应用于临床分离株的快速鉴定，方法 利用真菌保守区核糖体基因和可变区内转录间隔区基因为靶序列，应用生物素标记的通用引物进行念珠菌DNA的PCR扩增，将将该PCR产物分别与固定在微孔板上的都柏林念珠菌和白念珠菌特异性探针杂交，经酶联显色反应测其A值。结果 能特异地鉴别白念珠菌和都柏林念珠菌标准菌株，对108株常规培养方法从临床标本分离的白念珠菌检测。结果显示106株菌株仅白念珠菌探针检测阳性，另2株菌株仅都柏林念珠菌探针阳性，结论 微孔板双探针杂交法能快速，特异地鉴别都柏林念珠菌和白念珠菌。

Rapid Identification of Candida albicans and Candida dubliniensis Isolates With Two Specific Probes Simultaneously

Objective To develop a microtitration plate en zyme immunoassay to differentiate Candida albicans from Candida dubliniensis isolates using two specific probes s imultaneously.Methods The fun-gus-specific universal primers derived from the internal transcribed s pacer region of fungal rDNA were labeled with biotin,while the C.albicans or C.dubliniensis specific capture probes were coated on the microplates.Genomic DNA purified from the two species was amplified by PCR.The biotinylated p roducts were captured by the probes coated on the microplates.The A 405 value was finally determined by the c olorimetric assay.Results The two species of Candida could be detected specifically.Out of 108clinical isolates originally identified as C.albicans on the basis of germtube formation,two isolates were positive for C.dubliniensis and negative for C.albicans.The other106isolates were positive for C.albicans and negative for C.dubliniensis.Conclusions Two-specific-probe hybridization method is rapid and re liable for differentiating C.albicans from C.dubliniensis.