Simultaneous identification of Aspergillus and Mucorales species by reverse line blot(RLB) hybridization assay

Abstract

Abstract: Objective To Rapidly differentiate Aspergillus and Mucorales species by reverse line blot hybridization(RLB).Methods Totally 98 isolates including five Aspergillus strains(Aspergillus fumigatus,Aspergillus flavus,Aspergillus niger,Aspergillus terreus and Aspergillus nidulans) and seven Mucorales species(Mucor heimalis,Mucor racemosus,Mucor cercinelloidea,Rhizopus arrhizus,Rhizopus microsporus,Rhizomucor pusillus and Absidia corymbifera) were obtained from Research Center for Medical Mycology and Mycoses,Peking University.ITS1 and ITS4 fungal universal primers were chosen for PCR amplification,and the amplified products were used for reverse line blot hybridization with 12 fungal specie-specific probes.RLB data were compared with those by traditional fungal morphology and ITS sequencing methods.Results RLB showed high sensitivity and specificity,with 100% correct identification percentage of all the isolates and no cross hybridization between the species-specific probes.Eight negative control stains(Candida albicans,Fusarium solani,Scedosporium apiospermum,Penicillium marneffei,Exophiala verrucosa,Aspergillus clavatus,Aspergillus japonicus and Cunninghamella elegans) also showed negative by RLB.The analytical sensitivity of RLB was 1.8×10^-3 ng/μL by 10-fold serial dilution of Aspergillus fumigatus genomic DNA.Conclusions The RLB assay provides a rapid and reliable option for laboratory diagnosis and identification of Aspergillus and Mucorales species.

Key words: Aspergillus, Mucorales, PCR, Reverse line blot(RLB) hybridization

CLC Number:

ZHAO Zuo-tao, LI Li-li, WANG Xiao-yang, WAN Zhe, CHEN Wei, LI Ruo-yu. Simultaneous identification of Aspergillus and Mucorales species by reverse line blot(RLB) hybridization assay[J]. Chinese Journal of Mycology, 2011, 6(5): 261-266.

References

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