6株枝孢霉的鉴定与分析

Identification and analysis of six strains of Cladosporium

目的利用ITS基因序列分析对临床分离的6株枝孢霉进行分子生物学鉴定与分析,以提高对枝孢霉的鉴定能力。方法从皮肤病患者中分离6株枝孢霉,通过观察培养后菌落的形态及其显微镜下特征,并利用ITS序列对其进行PCR分子生物学分析,并将PCR产物送公司进行测序。结果 6株枝孢霉由显微镜下的形态和培养的菌落特征鉴定,提取其DNA进行PCR扩增,产物进行1%琼脂糖凝胶电泳分析,再由Tanon-3500凝胶成像系统确定扩增所得电泳带的大小在500~750 bp之间,测序结果经Blast比对,其中4株鉴定为球孢枝孢霉100%,1株鉴定为枝孢样枝孢霉100%,1株鉴定为枝孢霉属97%。结论通过ITS基因序列分析技术,能更有效准确地提高对枝孢霉的鉴定能力,为临床的准确治疗方案提供有力的依据。

Objective To perform molecular biological identification and analysis of six strains of Cladospori-um clinically by ITS gene sequencing, and to improve the ability to identify Cladosporium. Methods Six strains of Cladosporium were isolated from the patients with dermatosis. Colony characteristics and microscopic morphology of the isolates were observed by methods of culture and direct microscopy. PCR analysis of the isolates was performed by ITS gene sequencing and the PCR product was sent for sequencing. Results Six strains of Cladosporium could be identified by microscopic morphology and colony characteristics. DNA was extracted for PCR analysis, and PCR product was ana-lyzed on 1% agarose gel electrophoresis. The size of product was 500-750 bp using Tanon-3500 gel imaging system. By BLAST, four isolates displayed 100% consistence with Beauveria Cladosporium, one showed 100% consistence with cladosporium cladosporioides, and one had 97% consistence with Cladosporium. Conclusion ITS gene sequencing can identify Cladosporium effectively and provide strong basis for clinical treatment.