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羊呼吸道主要支原体和细菌病原的多重PCR检测方法的建立及应用 预览

Development and application of a multiplex PCR for major Mycoplasmas and bacterial respiratory pathogens of sheep and goats
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摘要 为建立一种能够同时检测绵羊肺炎支原体、丝状支原体簇成员、多杀性巴氏杆菌和溶血性曼氏杆菌4种羊呼吸道主要病原的多重PCR方法,本研究采用各病原的特异性引物,经优化反应条件,建立了一种可以同时检测以上4种病原的多重PCR方法,并对128份临床样品进行了检测。结果显示,该方法对其它细菌和支原体扩增结果均为阴性,特异性强;该方法对以上4种病原基因组DNA的检测下限分别为2.2×10^3拷贝、2.1×10^3拷贝、1.1×10^3拷贝和8.3×10^2拷贝。该方法特异性强、敏感性高,为上述4种病原的快速检测和鉴定,以及临床羊呼吸道疾病的诊断提供了一种方便、快捷和准确的工具。 To establish a method for simultaneous detection of four major Mycoplasmas and bacterial respiratory pathogens of sheep and goats, i.e.M.ovipneumoniae, M.mycoides cluster, Pasteurellamultocida and M.haemolytica, the multiplex PCR was developed with four pairs of specific primers designed for these bacteria, and the PCR cycle profile and components were optimized. Subsequently, the specificity and sensitivity of the assay were determined. A total of 128 clinical samples were also tested to validate the developed method. The results showed that the multiplex PCR was specific, sensitive and feasible for detection of clinical samples. The multiplex PCR established in this study would be useful for rapid detection of pathogens above and diagnosis of respiratory diseases in sheep and goats.
作者 胡玉婷 刀筱芳 王成龙 丁可莉 宋明彤 杨发龙 HU Yu-ting, DAO Xiao-fang, WANG Cheng-long, Ding Ke-li, Song Ming-tong, YANG Fa-long (College of Life Science and Technology, Southwest University for Nationalities, Chengdu 610041, China)
出处 《中国预防兽医学报》 CSCD 北大核心 2018年第5期416-420,430共6页 Chinese Journal of Preventive Veterinary Medicine
基金 国家重点研究计划(2016YFD0500907) 中央高校基本科研业务费专项资金项目(2015NZYQN61) 西南民族大学大学生创新创业训练计划项目(S201610656054)
关键词 绵羊肺炎支原体 丝状支原体簇 多杀性巴氏杆菌 溶血性曼氏杆菌 多重PCR Mycoplasma ovipneumoniae Mycoplasma mycoides cluster Pasteurellamultocida Mannheimiahaemolytica multiplex PCR
作者简介 胡玉婷(1995-),女,陕西咸阳人,本科,主要从事动物病原微生物学研究.;通信作者:E-mail:yfalong@hotmail.com
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