复合PCR检测医学常见重要真菌的研究

目的　建立复合PCR用于快速检测白念珠菌和非白念珠菌医学真菌。方法　设计白念珠菌特异引物用于扩增白念珠菌DNA ;11种医学真菌 (白念珠菌、热带念珠菌、假热带念珠菌、近平滑念珠菌、光滑念珠菌、解脂念珠菌、克鲁斯念珠菌、季也蒙念珠菌、新型隐球菌、烟曲霉和黄曲霉 )的DNA用医学真菌通用引物扩增 ;建立由上述两组引物组成的复合PCR ,在一次试验中区分白念珠菌和非白念珠菌真菌。结果　白念珠菌引物只特异扩增白念珠菌DNA ,扩增产物为 779bp ;通用引物扩增上述 11种医学真菌DNA ,扩增产物为 2 6 0bp左右。 12种常见致病细菌、乙肝病毒及人血液白细胞和组织细胞DNA均不被这两种引物扩增。复合PCR扩增白念珠菌DNA产生两条DNA片段 ,其长度分别为 779bp和 2 5 8bp ;非白念珠菌DNA只产生一条 2 6 0bp左右的片段。结论　复合PCR可在一次试验中检出白念珠菌和非白念珠菌 ,因此 ,复合PCR对于医学真菌的检测是一种较快速、敏感和特异的方法。

Study on the detection of the medical important fungi by multiple PCR

Aim To establish multiple PCR method for rapid detection of Candida albicans and non-Candida albicans medical fungi in one test.Method Designed primers of C.Albicans was used to amplify C.Albicans DNA.The DNA of 11species of fungi(C.Albicans,C.Tropicalis,C.Pseudotropicalis,C.Parapsilosis,C.Glabrata,C.Lipolytica,C.Krusi,C.Guilliermondii,Cryptococcus neoformans,Aspergillus fumigatus,A flavus.)were amplified by PCR using the universal primers of medical fungi.A multiple PCR,consisting of the two sets of primers was developed to allow in one test differentiating C.Albicans and other medical fungi.Results The DNA of C.Albicans was only amplified by PCR using specific primers of C.Albicans,while the DNA of other medical fungi were not amplified.The length of amplified fragment was 779bp.The DNA of fungi mentioned above were amplified by PCR using the universal primers.The DNA from 12 species of bacteria,hepatitis B virus and human WBC were not amplified by the two sets of primers.Two DNA fragments,779bp and 258bp in C.Albicans were generated using mixtures of two sets of primers,while 260bp DNA fragment in non-C.Albicans fungi was only generated.Conclusion Multiple PCR may rapidly detect C.Albicans and non-C.Albicans medical fungi in one test.It is a more rapid,specific and sensitive method for the detection of C.Albicans and non-C.Albicans medical fungi.