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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 To investigate the feasibility of typing the largest exon sequence of CAP5 9 gene in Cryptococcus neoformans. Methods According to the largest exon sequence of CAP5 9 gene, one pair of primers was designed and five serotypes of Cryptococcus neoformans were amplified by PCR. Sequences were analyzed by GenBank database. Phylip3.6a3 software was used to process the data and phylogenetic tree Tree, select a certain number of C. neoformans clinical isolates and other pathogenic fungi for clinical validation. Results The highest exon sequence of CAP5 9 in different serotypes of C. neoformans was different. Clinical validation showed that the largest exon of CAP5 9 gene conformed to the typing results of immunized typing. Conclusion The genes amplified by the designed primers can be used as molecular biology tools for typing and detecting Cryptococcus neoformans.