CAP59荚膜相关基因最大外显子序列用于新生隐球菌分型的探讨

The longest exon of CAP59 gene of C.neoformans and its application in detecting and typing C.neoformans

目的 探讨CAP5 9基因最大外显子序列对新生隐球菌进行分型的可行性。方法 根据CAP5 9基因最大外显子序列设计 1对引物 ,对新生隐球菌 5个血清型菌株进行PCR扩增和测序 ,用GenBank数据库分析序列的差别 ,Phylip3.6a3软件处理数据并绘制种系进化树 ,选择一定数量的新生隐球菌临床分离株和其他致病真菌进行临床验证。结果 新生隐球菌不同血清型菌株的CAP5 9最大外显子序列存在差别 ,临床验证显示CAP5 9基因最大外显子对受试菌株分型结果与免疫学分型结果全部符合。结论 以所设计引物扩增的基因可以作为新生隐球菌分型和检测的分子生物学工具。

Objective Potential application of the longest exon of CAP59 gene in the detection and typing of C.neoformans. Methods Primers were designed and the fragment was amplied according to the longest exon of CAP59 gene. The sequence of five serotypes of standard C.neoformans was analysed one by one in GenBank. Their phylogenetic tree was analysed with Phylip3.6a3. Some seotyped isolates of C.neoformans and other fugi were collected to test primers′s usefullness in detecting and seotyping of C.neoformans. Results There are differences of the longest exon of CAP59 gene among five serotypes of standard C.neoformans isolates. The serotying result of clinical isolates with the longest exon of CAP59 gene was all consistent with immunological serotyping. Conclusion The gene amplied by primers, P1: 5′-GAG TGT CTC CGC AAC CCG CA-3′ and P2: 5′-CCT ACT CTG CCA AAT CAA CTC-3′ is able to type and detect C.neoformans as a molecular too.