DNA扩增制备探针并用于临床白色念珠菌感染鉴定

目的应用聚合酶链反应技术扩增特异DNA片段,并制备成为可应用于临床鉴定白色念珠菌的基因探针. 方法采用聚合酶链反应技术特异地扩增白色念珠菌标准株DNA EO3 125 bp基因片段,然后用半抗原地高辛配基标记,在完成基因探针显色敏感度检测之后,应用该基因探针分别对血液病病房分离的白色念珠菌、热带念珠菌及大肠埃希菌等多种细菌进行斑点杂交测试. 结果基因扩增制备Dig-125 bp DNA探针显色敏感度为0.01 pg,具有白色念珠菌特异性,与其他念珠菌和细菌不发生杂交. 结论基因扩增制备EO3 125 bp DNA探针方法简便可靠,具有很强特异性,可应用于临床鉴定白色念珠菌感染.

Molecular Probe for Identification of Candida albicans Prepared by DNA Amplification

OBJECTIVE To prepare the special molecular probe by DNA amplification for identification of Candida albicans . METHODS The 125 bp DNA from EO 3 gene of C. albicans was amplified by PCR, then labeled by hepten digoxigenin. The sensitivity of special probe was detected, and the specificity was detected by dot blotting hybridization assay with 23 strains of C. albicans, C. parapsilosis, C. tropicalis and Escherichia coli etc isolated from patients of hematological department. RESULTS The Dig 125bp DNA probe prepared by DNA amplification only hybridized with C. albicans, whose sensitivity of color development was 0 01 pg. CONCLUSIONS The Dig 125bp molecular probe prepared by PCR from EO 3 DNA is specific and sensitive to C. albicans and useful for the identification of clinical C. albicans infection.