Amplified fragment length polymorphism analysis of Cryptococcus neoformans

Abstract

Abstract: Objective To evaluate AFLP-DNA fingerprinting method in the discrimination of C.neoformans.Methods Genomic DNA from C.neoformans was digested simultaneously with two restriction enzymes.Double-stranded adapters were ligated to the ends of restricted fragments and DNA fragments were amplified by PCR using primers complementary to the adapters and the restriction site sequences.AFLP products were separated using high resolution denaturing polyacrylamide gel electrophoresis and visualized by silver staining.Results DNA fingerprint patterns of 18 C.neoformans isolates from five serotypes and clinical isolates were analyzed by AFLP.AFLP fingerprints comprised over 30 bands detected in size range 30-500 bp.Different AFLP patterns were detected in isolates of the same serotype and identical banding patterns were observed from in isolates from the same patient at different time interals and different sources from one patiant.Conclusions AFLP is highly discriminatory,and powerful epidemiological tool for improving our understanding of C.neoformans.

Key words: Cryptococcus neoformans, DNA, AFLP

CLC Number:

R379.5

ZOU Xian-biao, WEN Hai, LIAO Wan-qing, WU Jian-hua, QIU Yun, YANG Jin-shui. Amplified fragment length polymorphism analysis of Cryptococcus neoformans[J]. Chinese Journal of Mycology, 2010, 5(2): 101-104.

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