Molecular identification and genotyping and in vitro drug sensitivity study of the first five clinical isolates of Cryptococcus neoformans in Xinjiang

Abstract

Abstract:

Objective For the first time to perform the molecular identification,RAPD-PCR genotyping and in vitro drug sensitivity study of 5 clinical strains of Cryptococcus neoformans in Xinjiang.Methods Molecular classification using primer SEQ-6 from big subunit ribosomal DNA (LSU rDNA) D1/D2 gene region and RAPD-PCR method were applied in 5 strains of clinical isolated Xinjiang Cryptococcus neoformans strains and 5 Cryptococcus strains provided by Shanghai Changzheng hospital.Sensitivity in vitro of 5 clinical isolated Xinjiang Cryptococcus neoformans strains of the above 6 kinds of antifungal drugs (Voriconazole,itraconazole,amphotericin B,terbinafine,fluconazole,5-fluorine cytosine)were judged according to clinical laboratory standards institute (CLSI) trace liquid dilution method of yeast (M27-A3).Results The 5 strains of Xinjiang were identified as Cryptococcus neoformans through D1/D2 region sequence analysis.All 10 strains are divided into A,B,C,D 4 genotype by RAPD-PCR.One Xinjiang strain was type A,the rest 4 strains were type B;Among 5 Shanghai strains,1 strain was type B,3 strainswere type C,1 strain was typeD.For the 5 Xinjiang clinical Cryptococcus neoformans strains,to the MIC value ranges of voriconazole,itraconazole,amphotericin B and terbinafine were as following:0.062 5-0.25 μg/mL,0.25-1 μg/mL,0.125-0.5 μg/mL,1-2 μg/mL,respectively;MIC value ranges of fluconazole,5-fluorine cytosine MIC value were much higher as:4-16 μg/mL,8-32 μg/mL,respectively.MIC values of genotype A and B to voriconazole were 0.062 5-0.125 μg/mL.Conclusion The 5 Xinjiang clinical isolates of Cryptococcus were identified as Cryptococcus neoformans using D1/D2 genes analysis.RAPD-PCR revealed these clinical Cryptococcus neoformans strain in Xinjiang with priority given to type B genotypes.Type A and type B isolates weremore sensitive to voriconazole,amphotericin B,itraconazole and terbinafine.

Key words: Cryptococcus neoformans, Xinjiang, RAPD-PCR genotyping, antifungal sensitivity

CLC Number:

R379.5

Yultuz·Xohrat, Kadirye·Kasim, WANG Xiao-dong, DENG Shu-wen, Prida·Abliz. Molecular identification and genotyping and in vitro drug sensitivity study of the first five clinical isolates of Cryptococcus neoformans in Xinjiang[J]. Chinese Journal of Mycology, 2016, 11(6): 327-331.

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