应用RAPD技术鉴定地霉的实验研究

目的探讨RAPD技术在快速鉴定地霉中应用。方法用E.Z.N.A.yeastDNAkit提取地霉菌基因组DNA,采用随机引物AP3(5'-TCGTAGCCAA-3')、ATG(5'-ATGGATCGGC-3')、RP2(5'-AAGGATCAGA-3')、OPA-10(5'-GTGATCGCAG-3')对临床上致病性白地霉、林生地霉皮损株和血液株的基因组DNA进行扩增,对各病原菌的DNA指纹的特征进行分析。结果成功提取了地霉的基因组DNA,其纯度和浓度均能满足PCR反应的要求。利用4种引物对基因组DNA进行扩增,不同种真菌的DNA显示不同的DNA带型,分离自不同感染部位的同种不同株真菌的DNA显示的主要DNA带型基本一致。结论采用E.Z.N.A.yeastDNAkit提取的地霉基因组DNA可以用于PCR反应。RAPD法鉴定地霉菌简单、快速、特异,可用于临床诊断。

Identification of Geotrichum by RAPD

Objective To study the random amplified polymorphic DNA for rapid identification of Geotrichum.Methods DNAs of Geotrichum candidum,Geotrichum silvicola isolated from the skin lesion and blood were extracted by E.Z.N.A.yeast DNA kit and amplified using AP3（5＇-TCGTAGCCAA-3＇）,ATG（5＇-ATGGATCGGC-3＇）,RP2（5＇-AAGGATCAGA-3＇）,OPA-10（5＇-GTGATCGCAG-3＇）as primers.The characteristics of fingerprint was analyzed.Results DNA of Geotrichum was successfully extracted with satisfied purity and concentration for PCR.Distinct DNA band patterns showed differences in different Geotrichum species while displayed similarity within different strains of same species isolated from different lesions.Conclusions DNA extracted by E.Z.N.A.yeast DNA kit could applied for PCR.RAPD method is specific,rapid and simple for Geotrichum indetification.