微卫星引物PCR快速鉴定兔皮肤病原真菌方法的建立及应用

畜牧兽医学报 ›› 2012, Vol. 43 ›› Issue (5): 779-784.doi:

微卫星引物PCR快速鉴定兔皮肤病原真菌方法的建立及应用

李代军1，周玉法2，刘敬博1，高丽丽1，赵效楠1, 蔡玉梅1*，柴同杰1*

1.山东农业大学动物科技学院，泰安 271000； 2.泰安市岱岳区畜牧兽医局，泰安 271000

收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2012-05-28 发布日期:2012-05-28
通讯作者: 蔡玉梅1*，柴同杰1*

Development and Application of Microsatelliteprimered PCR for Rapidly Identifying Dermatophytes from Rabbits

LI Daijun1, ZHOU Yufa2, LIU Jingbo1, GAO Lili1, ZHAO Xiaonan1,
CAI Yumei1*, CHAI Tongjie1*

1. College of Animal Sciences, Shandong Agricultural University, Tai’an 271000, China;2. Animal Husbandry Bureau of Daiyue Region, Tai’an 271000, China

Received:1900-01-01 Revised:1900-01-01 Online:2012-05-28 Published:2012-05-28
Contact: CAI Yumei1*, CHAI Tongjie1*

摘要/Abstract

摘要： 本研究旨在借助微卫星引物PCR和聚类分析软件，建立一种快速准确鉴定兔皮肤病原真菌的分子生物学方法。本试验以寡核苷酸重复序列(GACA)4为引物，首先对3种常见兔皮肤病原真菌（须癣毛癣菌、犬小孢子菌和石膏样小孢子菌）标准菌株的DNA进行扩增，然后用NTSYSpc2.10软件分析其DNA指纹的多态性，并根据其多态性的差异进行鉴定分析。最后，用上述方法对临床上的分离株进行鉴定分析，并与传统的形态学鉴定方法进行比较。结果表明，3种不同的兔皮肤病原真菌均呈现不同的DNA指纹，且条带差异明显；聚类分析图显示，同种病原真菌的DNA指纹有很高的相似性（90%~100%）；该方法对分离株的鉴定结果与形态学鉴定结果一致。可见，通过微卫星引物PCR结合聚类分析软件可以准确、快速地鉴定兔皮肤病原真菌。

Abstract: The study was carried out to establish an accurate and rapid method to identify dermatophytes from rabbits by microsatelliteprimered polymerase chain reaction (PCR) and NTSYSpc2.10 software. The DNAs of Trichophyton mentagrophyton, Microsporum gypseum and Microsporum canis were amplified by the primer (GACA)4, and then their DNA fingerprint polymorphism was analyzed using NTSYSpc2.10 software. Based on the above results, the identification method was confirmed. Finally, a total of 21 clinical isolates identified by morphology were identified by the method. The results showed that: 1) The three kinds of rabbit dermatophytes have obviously different DNA fingerprints; 2) The rabbit dermatophytes of the same kind have high genetic similarity (90%100%); 3) The identified results of clinical isolates by this method were consistent with the morphological identification results. The results indicated that dermatophytes from rabbits could be rapidly and accurately identified by the microsatelliteprimered PCR in combination with NTSYSpc2.10 software.

李代军;周玉法;刘敬博;高丽丽;赵效楠;蔡玉梅;柴同杰. 微卫星引物PCR快速鉴定兔皮肤病原真菌方法的建立及应用[J]. 畜牧兽医学报, 2012, 43(5): 779-784.

LI Daijun;ZHOU Yufa;LIU Jingbo;GAO Lili;ZHAO Xiaonan;CAI Yumei;CHAI Tongjie. Development and Application of Microsatelliteprimered PCR for Rapidly Identifying Dermatophytes from Rabbits[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2012, 43(5): 779-784.

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