Abstract: Objective To develop a PCR method to detect common deep pathogenic fungi for the application in clinical settings.Methods A hot-start polymerase chain reaction (PCR)-based method was developed.A simplex PCR with a universal fungus-specific primer set was used to detect experimental and simulating clinical specimens.For specimens with positive results,a triplex PCR was further applied with the species-specific primers of Candida albicans,Aspergillus fumigatus and Cryptococcus neoformans.Results A 260 bp product was successfully amplified from all 78 strains of 55 fungal species in 9 fungal genera,but not amplified from any strain of other microbes and human cells.The detection system was of high specificity and sensitivity,and convenient and inexpensive for use.Conclusion The simplex and triplex PCR approach based on highly specific hot-start PCR might be useful for quick detection of deep fugal infection in routine clinical laboratory practice.

Key words: Candida, Aspergillus, Cryptococcus, Polymerase chain reaction