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| 应用TaqMan荧光定量PCR检测牛分枝杆菌 | |
| 引用本文： | 王春雨,王振国,刘金华,宋战昀,周亮,高宏伟,王全凯. 应用TaqMan荧光定量PCR检测牛分枝杆菌[J]. 中国预防兽医学报, 2011, 33(2). DOI: 10.3969/j.issn.1008-0589.2011.02.13 |
| 作者姓名： | 王春雨 王振国 刘金华 宋战昀 周亮 高宏伟 王全凯 |
| 作者单位： | 1. 吉林农业大学,吉林,长春,130118 2. 吉林出入境检验检疫局检验检疫技术中心,吉林,长春,130062 3. 军事医学科学院军事兽医研究所,吉林,长春,13006 |
| 基金项目： | 吉林省科技发展计划国际合作项目(20100718) |
| 摘 要： | 为建立快速检测牛分枝杆菌(M.bovis)的TaqMan荧光定量PCR方法,本研究以GenBank登录的M.bovis特有229 bp基因为研究对象,设计并合成引物及探针。该方法具有较好的特异性,与标准质控菌株呈阳性反应,与其他微生物样品呈阴性反应;灵敏性最低检测值可达1 pg/mL;对20阳性临床样品进行荧光定量PCR检测,均为阳性;而对培养为阴性的20份临床样品进行检测,6份为阳性。该研究结果表明,建立的方法特异性强,敏感性高,稳定性好,能够用于M.bovis的鉴别检测,对牛分枝杆菌病的快速检测和早期诊断具有重要意义。 |
| 关 键 词： | 牛分枝杆菌病 鉴别检测 牛型结核杆菌 荧光定量PCR |
Detection of Mycobacterium bovis with TaqMan real-time PCR |
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| WANG Chun-yu,WANG Zhen-guo,LIU Jin-hua,SONG Zhan-yun,ZHOU Liang,GAO Hong-wei,WANG Quan-kai. Detection of Mycobacterium bovis with TaqMan real-time PCR[J]. Chinese Journal of Preventive Veterinary Medicine, 2011, 33(2). DOI: 10.3969/j.issn.1008-0589.2011.02.13 | |
| Authors: | WANG Chun-yu WANG Zhen-guo LIU Jin-hua SONG Zhan-yun ZHOU Liang GAO Hong-wei WANG Quan-kai |
| Affiliation: | WANG Chun-yu1,WANG Zhen-guo2,LIU Jin-hua2,SONG Zhan-yun2,ZHOU Liang1,GAO Hong-wei3*,WANG Quan-kai1*(1.Jilin Agricultural University,Changchun 130118,China,2.Jilin Entry-Exit Inspection and Quarantine Bureau,Changchun 130062,3.Research Institute of Veterinary,Academy of Military Medical Sciences,China) |
| Abstract: | A TaqMan real-time PCR assay was developed for detection of Mycobacterium bovis infection in cattle based on primers and TaqMan probe derived from M.bovis sequence in GenBank.The specific test showed that the assay had positive results for detection of M.bovis strains and negative for other bacteria.This assay could detect single bacteria.Comparing with other methed on 20 PPD positive clinical samples which were negative by bacteria isolation,six samples were positive detected by the real-time PCR.Indicatin... |
| Keywords: | bovine tuberculosis specific detection Mycobacterium bovis real-time PCR |
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