Abstract: Objective To evaluate the status of Cryptococcus neoformans var.shanghaiensis S8012 and assess sequence diversity of ITS region among C.neoformans varieties by sequence analysis and phylogenesis analysis on this region.Methods Universal primers(ITS1 and ITS4) were designed and used to amplify the ITS region(including 5.8S rRNA gene) of 12 reference strains of C neoformans varieties by PCR,and the products were sequenced.The resultant sequence was blasted against the sequences of 11 reference strains of C.neoformans by CLUSTAL W1.83 software with several databases including Genebank.MEGA3.1 software was applied to process data and draw the phylogenetic tree.Results There were significant differences in the sequence of ITS region among C.neoformans varieties.As shown by the sequence analysis of ITS region,six subtypes(A-F) were identified in the three varieties of C.neofonnans and 4 subtypes(C,D,E and F) in C.neofonnans var.gattii.Significant differences were observed in the phenotype and molecular biology between C.neoformans var shanghaiensis 58012 and var.gattii,but the diversity was within species variation.Conclusions The C.neoformans var.shanghaiensis 58012 isolated from Shanghai,China,belongs to subtype C of C.neoformans var.gattii based on ITS sequence analysis,and the original C.neoformans var.gattii RV20186 belongs to subtype F of C.neoformans var.gattii.

Key words: Cryptococcus neofonnans, Variety, Sequence analysis,ITS