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猪肺炎支原体和猪鼻支原体双重PCR检测方法的建立与应用
刘茂军,邵国青
0
(江苏省农业科学院兽医研究所?农业部兽用生物制品工程技术重点实验室?国家兽用生物制品工程技术研究中心;南京天邦生物科技有限公司)
摘要:
根据猪肺炎支原体(Mhp)和猪鼻支原体(Mhr)的16S rRNA基因设计3条引物, 建立Mhp和Mhr的双重PCR检测方法,并对该方法进行了特异性和敏感性试验,并使用建立的方法检测了临床样品和疫苗样品。结果显示该方法具有良好的特异性,最低可检测到0.66ng 的Mhp基因组DNA和0.58 ng Mhr基因组DNA,临床样品和疫苗样品检测结果与普通PCR检测结果一致。该双重PCR方法,可用于Mhp与Mhr的鉴别、诊断以及疫苗纯粹性检查,快速而准确。
关键词:  猪支原体肺炎  猪鼻支原体  PCR  检测
DOI:
投稿时间:2012-06-27修订日期:2012-07-12
基金项目:国家自然科学基金(31100136)、江苏省农业科技自主创新资金项目[CX(11)3025]
Establishment and application of Double PCR method for detection of Mycoplasma hyopneumoniae and Mycoplasma hyorhinis
(Institute of Veterinary Medicine,Jiangsu Academy of Agricultural Sciences Key Laboratory of Veterinary Biological Engineering and Technology,Ministry of Agriculture of the P R China National Research Center for Engineering and Technology of Veterinary Biological Products,Nanjing,;Nanjing Tianbang Bio-Industry Co Ltd)
Abstract:
According to the 16S rRNA gene of Mhp and Mhr, we designed three primers and established a double PCR method for simultaneous detectting Mycoplasma hyopneumoniae(Mhp) and Mycoplasma hyorhinis(Mhr), and the specificity and sensibility assay were performed. The clinical samples and vaccine sample were detected. The result showed the gene could be specifically amplified from Mhp and Mhr. In addition, the method could detect at least 0.66 ng of Mhp genome DNA and 0.58 ng of Mhr genome DNA. The results of detecting clinical samples and vaccine samples tested with double PCR were consisted with the normal PCR test that detecting Mhp by PCR established on P36 geng and Mhr on P37 gene. Therefore, the double PCR method is a fast and accurate method for detecting and diagnosing of Mhp and Mhr.
Key words:  Mycoplasma hyopneumoniae  Mycoplasma hyorhinis  PCR  Detection

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