Leachii支原体PCR检测方法的建立与应用

发布时间：2018-05-11 13:41

本文选题：leachii支原体 + PCR　；参考：《中国预防兽医学报》2017年11期

【摘要】：为建立一种leachii支原体的检测方法,本研究根据leachii支原体LPPA外膜蛋白编码基因序列设计一对引物,建立了PCR检测方法。结果显示,该方法仅对leachii支原体扩增出539 bp的目的片段,而与牛支原体、丝状支原体山羊亚种、绵羊肺炎支原体、丝状支原体丝状亚种小菌落型、山羊支原体山羊肺炎亚种等其它支原体无交叉反应,具有良好的特异性;该方法对leachii支原体的最低检测浓度为105 ccu/m L,敏感性较高。采用该方法可以在乳汁和关节液等病料样品中检测到leachii支原体,与分离培养的结果相符。上述结果表明,本研究建立的leachii支原体PCR检测方法具有特异性强、敏感性高的特点,可实现准确、高效检测leachii支原体的目的。
[Abstract]:In order to establish a method for detection of Mycoplasma leachii, a pair of primers was designed based on the sequence of LPPA outer membrane protein encoding gene of Mycoplasma leachii, and a method of PCR detection was established. The results showed that the method only amplified 539 BP target fragments for Mycoplasma leachii, but with bovine mycoplasma, Mycoplasma filamentum subspecies, Mycoplasma sheeppneumoniae. The Mycoplasma filamentous subspecies of Mycoplasma filamentous subspecies, the Mycoplasma goat pneumonia subspecies and other Mycoplasma have no cross reaction and have good specificity. The minimum detection concentration of Mycoplasma leachii is 105 ccu/m L, and the sensitivity is high. This method can be used to detect Mycoplasma leachii in the samples of milk and joint fluid. The results showed that the PCR detection method of Mycoplasma leachii established in this study has the characteristics of high specificity and high sensitivity, which can be used for accurate and efficient detection of leachii mycoplasma.

【作者单位】：黑龙江八一农垦大学动物科技学院;中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室/牛羊传染病研究创新团队;
【基金】：黑龙江省自然科学基金面上项目(C201348) 兽医生物技术国家重点实验室开放课题基金(sklvbf201604)
【分类号】：S852.62

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