摘要
本试验旨在建立一种可直接从病料中快速检测牛支原体的PCR方法,并且应用于临床。根据牛支原体16S rRNA基因设计一对引物,配制PCR扩增体系,样品经简单处理后加入体系进行扩增,最终扩增出1 390bp片段,进一步的试验证明该方法具有良好的特异性和敏感性。临床样品检测结果显示,该方法的检出率与传统PCR的符合率为100%。试验表明,该方法能快速直接从临床样品中检测出牛支原体,可用于牛支原体的大量临床样品的检测。
The aim of this study was to develop a rapid and direct PCR method to detect Mycoplasma bovis from diseased materials. A pair of primers were designed according to the sequence of 16 S rRNA gene of Mycoplasma bovis, and the system of direct PCR amplification was prepared. After simple treatment, the samples were added into the system for amplification and electrophoresis. A 1390 bp fragment was amplified, and the specificity and sensitivity were tested. The results showed that the method was specific and sensitive. The method was applied to detect clinical samples, and the detection rate was 100% conform with the traditional PCR. The method can directly detect Mycoplasma bovis from clinical samples and can be used for the detection of a large number of clinical samples of Mycoplasma bovis.
作者
徐引弟
李海利
王治方
张青娴
索延乐
宋振宇
XU Yin-di LI Hai-li WANG Zhi-fang ZHANG Qing-xian SUO Yan-le SONG Zhen-yu(Institute of Animal Husbandry and Veterinary Research, Henan Academy of Agricultural Sciences, Zhengzhou 450002 Henan Key Laboratory of Farm Animal Breeding and Nutritional Regulation, Zhengzhou 450002 Animal Health Inspection in Jiyuan City Henan Province, Jiyuan 459000)
出处
《中国奶牛》
2017年第6期41-44,共4页
China Dairy Cattle
基金
河南省科技攻关计划项目(162102110042)
河南省农业科学院自主创新基金项目(2016ZC51
2017ZC49)
关键词
快速直接PCR
牛支原体
特异性
敏感性
临床应用
Rapid and direct PCR method
Mycoplasma bovis
Specificity
Susceptibility
Clinical application
