PCR法扩增角鲨烯环氧化酶鉴定白念珠菌的研究

目的：通过扩增角鲨烯环氧化酶基因的开放读框，进行PCR来鉴定白念珠菌。方法：根据白念珠菌角鲨烯环氧化酶基因的开放读框中编码1MSSVKY^6的序列设计上游引物5‘－ATGAGTTCAGTTAAGTATG－3‘，编码^492NEIVR^496的序列设计下游引物5‘-CTATCTTACAATCTCGTTC-3‘，对白念珠菌ATCC11006，22株临床分离株，7株其它致病性念珠菌，8株致病性丝状真菌，1株新生隐球菌及1份人的基因组DNA进行了PCR扩增，并对PCR产物用酶切方法鉴定，结果：所有23株白念菌均可获得约1．5kb大小的PCR产物，16株其它致病性真菌及人的DNA标本均无扩增产物，PCR产物经BamHI酶切，产生两个大小分别约为1．0kb和500bp的片段。结论：利用设计的引物，扩增角鲨烯环氧化酶基因的开放读框，可以特异地鉴定白念珠菌。

Identification of Candida albicans by PCR amplification of the open reading frame coding squalene epoxidase

Objectives:To identify Candida albicans by PCR amplification of the open reading frame coding squalene epoxidase. Method:The primers, forward: 5' ATGAGTTCAGTTAAGTATG 3' and reverse: 5' CTATCTTACAATCTCGTTC 3',which contain the sequences corresponding to amino acid 1 MSSVKY 6 and 492 NEIVR 496 of squalene epoxidase from C.albicans, were synthesized. Polymerase chain reaction (PCR) was applied using these primers and genomic DNAs from C.albicans,other non C.albicans fungi and human genomic DNA as template. The PCR product of C.albicans ATCC11006 was identified by the digestion with BamHI.Results: About 1.5 kb products were obtained from all of the 23 C.albicans strains including 22 clinical isolates and ATCC11006. Two about 1.0 kb and 0.5 kb fragments were produced from the digestion of the PCR product of C.albicans ATCC11006 with BamHI. No product can be amplified from 16 strains of non C.albicans fungi and human genomic DNA. Conclusion : C.albicans can be identified specially by PCR amplification of the open reading frame coding squalene epoxidase.