| 猪支原体不同靶基因PCR检测方法的比较 |
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| 引用本文： | 刘明明,贾立军,薛书江,梁晚枫,张守发.猪支原体不同靶基因PCR检测方法的比较[J].畜牧与兽医,2012,44(3):1-4. |
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| 作者姓名： | 刘明明 贾立军 薛书江 梁晚枫 张守发 |
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| 作者单位： | 延边大学农学院动物医学系,吉林延吉,133002 |
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| 基金项目： | 公益性行业(农业)科研专项(200903036-13) |
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| 摘 要： | 为筛选出检测猪支原体更为特异、敏感的PCR检测方法,本试验分别以16S rRNA、50S rRNA和膜蛋白OxaA为靶基因进行PCR检测,并从其敏感性、特异性和临床样本检出率等方面进行了比较。结果显示,以膜蛋白OxaA和16S rRNA为靶基因的PCR方法敏感性最高,最小检测DNA量为1.86 fg/μL,而以50S rRNA为靶基因的PCR方法最小检测DNA量为18.6 fg/μL;3种靶基因引物均扩增不出大肠杆菌、猪链球菌、猪肺炎支原体、牛附红细胞体等基因片段,具有较好的特异性;通过对临床60份血液样本的检测结果表明,以膜蛋白OxaA基因设计的引物检出率最高,为25%(15/60),明显高于16S rRNA基因的21.6%(13/60)和50S rRNA基因的18.3%(11/60)。本试验为猪支原体病的诊断及流行病学调查提供了更为敏感、特异的检测技术。
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| 关 键 词： | 猪支原体 16S rRNA 50S rRNA 膜蛋白OxaA |
Comparison of PCR methods to different target genes of Mycoplasma suis |
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| LIU Ming-ming
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JIA Li-jun
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XUE Shu-jiang
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LIANG Wan-feng
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ZHANG Shou-fa.Comparison of PCR methods to different target genes of Mycoplasma suis[J].Animal Husbandry & Veterinary Medicine,2012,44(3):1-4. |
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| Authors: | LIU Ming-ming JIA Li-jun XUE Shu-jiang LIANG Wan-feng ZHANG Shou-fa |
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| Institution: | (Department of Veterinary Medicine,Agriculture College of Yanbian University,Yanji 133002,China) |
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| Abstract: | The aim of this study is to develop a s PCR method for the specific and sensitive identification of Mycoplasma suis.The genes encoding 16S rRNA,50S rRNA and membrane protein OxaA were used as detection targets.The results showed that PCR methods based on OxaA and 16S rRNA genes had higher sensitivity and the minimum detectable concentration was 1.86fg/μL.However,the minimum detectable concentration of 50S rRNA gene was 18.6fg/μL.The genes of E.coli,Streptococcus,M.hyopneumoniae and M.wenyonii could not be amplified by the above PCR methods.The detection results for 60 clinical samples showed that the PCR method based on OxaA gene(15/60) had significantly higher detection rate than that based on 16S rRNA gene(13/60) and 50S rRNA gene(11/60).The study provided a more sensitive and specific detection technique for diagnosis and epidemiological investigation of M.suis infection. |
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| Keywords: | Mycoplasma suis 16S rRNA 50S rRNA membrane protein OxaA |
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