[1]杨 帆,徐 娜,雷 宇,等.牛副流感3型病毒、牛传染性鼻气管炎病毒、牛病毒性腹泻病毒和牛支原体多重PCR检测方法的建立[J].中国预防兽医学报,2018,(05):411-415.[doi:10.3969/j.issn.1008-0589.201707011]
 YANG Fan,XU Na,LEI Yu,et al.Establishment of a multiplex PCR for detection of bovine parainfluenza virus 3, infectious bovine rhinotracheitis virus, bovine viral diarrhea virus and Mycoplasma bovis[J].Chinese journal of preventive veterinary medicine,2018,(05):411-415.[doi:10.3969/j.issn.1008-0589.201707011]
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牛副流感3型病毒、牛传染性鼻气管炎病毒、牛病毒性腹泻病毒和牛支原体多重PCR检测方法的建立()
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《中国预防兽医学报》[ISSN:1008-0589/CN:23-1417/S]

卷:
期数:
2018年05
页码:
411-415
栏目:
诊断和检测技术
出版日期:
2018-05-15

文章信息/Info

Title:
Establishment of a multiplex PCR for detection of bovine parainfluenza virus 3, infectious bovine rhinotracheitis virus, bovine viral diarrhea virus and Mycoplasma bovis
文章编号:
1008-0589(2018)5-0411-05
作者:
 

杨 帆12徐 娜12雷 宇12郝瑞峰3李平安12关平原12*

 (1. 内蒙古农业大学 兽医学院,内蒙古 呼和浩特 010018;2. 农业部动物疾病临床诊疗技术重点实验室,
内蒙古 呼和浩特,010018;3. 内蒙古自治区托克托县农产品质量安全检验检测站,内蒙古 托克托县 010200)
Author(s):
 

YANG Fan12 XU Na12 LEI Yu12 HAO Rui-feng3 LI Ping-an12 GUAN Ping-yuan12*

 (1. College of Veterinary Medicine, Inner Mongolia Agricultural University, Hohhot 010018, China;
2. Key Laboratory of Clinical Diagnosis and Treatment Technology in Animal Disease, Ministry of Agriculture, Hohhot 010018, China;
3. Inspection station for agricultural products quality and safety of Togtoh country in Inner Mongolia, Togtoh 010200, China)
关键词:
牛副流感3型病毒牛传染性鼻气管炎病毒牛病毒性腹泻病毒牛支原体多重PCR检测
Keywords:
  BPIV3  IBRV  BVDV  M.bovis  multiplex PCR  detection
分类号:
S852.65
DOI:
10.3969/j.issn.1008-0589.201707011
文献标志码:
A
摘要:
为建立检测牛副流感3型病毒(BPIV3)、牛传染性鼻气管炎病毒(IBRV)、牛病毒性腹泻病毒(BVDV)和牛支原体(M.bovis)的多重PCR检测方法,本研究根据GenBank中登录的BPIV3的HN基因、IBRV的gC基因、BVDV的5’UTR和M.bovis的oppD/F基因序列设计特异性引物,通过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。结果显示,该方法对牛呼吸道合胞体病毒、小反刍兽疫病毒、牛布鲁氏菌、羊布鲁氏菌、牛源多杀巴氏杆菌A型和B型无特异性扩增,对BPIV3和BVDV的cDNA最低检测量分别为100 pg和1 ng ,对IBRV和M.bovis的DNA最低检测量分别为10 pg和100 pg,具有较高的特异性和灵敏性。对临床33份鼻拭子样品和12份牛肺样品的检测结果与已发表文献中PCR方法的检测结果一致。本研究建立的多重PCR检测方法可同时对BPIV3、IBRV、BVDV和M.bovis进行检测,为这4种病原的诊断和检测提供了一种实用、便捷的方法。
 
Abstract:
To establish a detection method for simultaneously detecting bovine parainfluenza virus 3 (BPIV3),  infectious bovine rhinotracheitis virus (IBRV), bovine viral diarrhea virus (BVDV) and Mycoplasma bovis, the multiplex PCR (mPCR) assay was developed with a set of specific primers designed based on HN, gC, 5’UTR, oppD/F genes of BPIV3, IBRV, BVDV and M.bovis, respectively. Meanwhile, the primers ratio and annealing temperature were optimized. The detection results showed that the mPCR was able to simultaneously amplify the specific fragments of 510bp for BPIV3, 308bp for IBRV, 215bp for BVDV and 78bp for M.bovis, which had no any amplification for BRSV, PPRV, B.melitensis, B.abortus, bovine P.multocida serotype A and bovine P.multocida serotype B. The detection sensitivity of the mPCR for viral DNA/cDNA of BPIV3, IBRV, BVDV and M.bovi were 100 pg, 10 pg, 1 ng and 100 pg, respectively. Additionally, the established mPCR assay was compared with traditional single PCR for 33 nose swab samples and 12 bovine lung samples detections, and the coincidence rate was 100%. These results demonstrated that the established mPCR was high effectively and sensitivity in detecting the four pathogenic microorganisms.

参考文献/References:

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(本文编辑:李    爽)

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更新日期/Last Update: 2018-05-15