60株临床分离犬小孢子菌的聚合酶链反应-限制性片段长度多态性鉴定

Identification of 60 strains of clinical isolated Microsporum canis with restriction fragment length polymorphism

目的探索采用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)方法进行犬小孢子菌的快速鉴定。方法采用真菌通用引物第1内转录间隔区(ITS-1)与第4内转录间隔区(ITS-4)对60株犬小孢子菌进行PCR扩增,然后利用限制性内切酶TaqⅠ和HinfⅠ分别对PCR产物进行酶切分析。结果真菌通用引物ITS-1和ITS-4在60株犬小孢子菌中均扩增出1条长约750 bp的DNA条带;2种内切酶HinfⅠ和TaqⅠ各自显示相同的酶切结果,酶切片段稳定而清晰。结论应用PCR-RFLP方法可以快速、敏感、特异地鉴定犬小孢子菌。

Objective To investigate a rapid method for the identification of Microsporum canis with polymerase chain reaction （PCR） and restriction fragment length polymorphism （RFLP）. Methods 60 strains of isolated Microsporum canis were amplified by PCR with fungi-universal primers first internal transcribed spacer and fourth internal transcribed spacer. Then RFLP was performed by using 2 restriction enzymes （Hinf I and Taq I ） to digest the production of PCR amplification. Results The pattern of 750 bp was obtained from Microsporum canis after PCR amplification and the patterns were identical by restriction enzyme Hinf I or Taq I. Conclusion PCR-RFLP can rapidly, sensitively and specifically identify Microsporum canis.