Rapid Detection and Identification of Pathogenic Fungi of Some Deep Fungal Infections by PCR in Combination with Genescan Analysis / 中华皮肤科杂志

ABSTRACT

Objective To rapidly detect and identify pathogenic fungi of some deep fungal infections by PCR.Methods The suspensions of22pathogenic fungi(23strains)were amplified by PCR with fungal universal primers ITS86and ITS4which were labeled by FAM.The precise length of amplified fragments was determined by ABI PRISM TM 377Sequencer and Genescan analysis software,then compared with that of am-plicons of corresponding fungal DNA which were previously extracted.Results(1)Amplification of17pathogenic fungi with ITS4,ITS86resulted in a unique fragment length(except for A.nidulans and A.niger,C.albicans and C.stellatoidea,F.pedrosoi and E.dermatitidis).(2)No significant difference of the length of am-plicons was found between the fungal suspension and control organisms,based on the results of Genescan analysis.(3)The whole process took only6h to complete the detection.Conclusion The combination of fun-gal suspension PCR with ITS fungal universal primers and Genescan analysis might provide an accurate,spe-cific,sensitive,and rapid approach to detect and identify22pathogenic fungi causing deep fungal infections,and hold promise to be applied for the diagnosis of deep fungal infection.