Abstract:

Objective To investigate the determinations of Candida,Cryptococcus neoformans and ERG11 gene point mutations by gene chip.Methods The primers and oligonucleotide probes of 5.8 S rDNA and 28S rDNA of internal transcribed spacer-2（ITS-2） gene of 5 species of pathogenic Candida and Cryptococcus neoformans were synthesized,and the oligonucleotide microarray was prepared for identification. The specific primers and oligonucleotide probes of point mutations in ERG11 gene were synthesized,and ERG11 gene was amplified by asymmetric fluorescence polymerase chain reaction（PCR）. The products were analyzed by oligonucleotide microarray.Results A total of 4 point mutations of ERG11 gene were amplified from 12 isolates of drug resistant Candida by asymmetric fluorescence PCR with specific primers,and 5 species of pathogenic Candida and Cryptococcus neoformans were identified by gene chip successfully.Conclusions The oligonucleotide microarray established in this study can be used to identify the clinical isolated pathogenic Candida and Cryptococcus neoformans.

Key words: Gene chip, Asymmetric fluorescence polymerase chain reaction, Pathogenic Candida, Cryptococcus neoformans