中国乳业 ›› 2023, Vol. 0 ›› Issue (7): 30-34.doi: 10.12377/1671-4393.23.07.07

• 奶牛传染病专题 • 上一篇    下一篇

交叉引物扩增法快速检测牛病毒性腹泻病毒

贺泂杰   

  1. 中国农业科学院兰州畜牧与兽药研究所,甘肃省中兽药工程技术研究中心,甘肃兰州 730050
  • 出版日期:2023-07-25 发布日期:2023-08-07
  • 作者简介:贺泂杰（1987-）,男,陕西榆林人,博士,副研究员,研究方向为预防兽医学及天然药物。
  • 基金资助:
    甘肃省科技计划项目-重点研发计划（21YF5NA143）

Rapid Detection of BVDV by Cross-priming Amplifcation

HE Jiongjie   

  1. Lanzhou Institute of Husbandry and Pharmaceutical Sciences,Chinese Academy of Agricultural Sciences,Engineering Technology Research Center of Traditional Chinese Veterinary Medicine of Gansu Province,Lanzhou Gansu 730050
  • Online:2023-07-25 Published:2023-08-07

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摘要： [目的]牛病毒性腹泻病毒（BVDV）是一种可以引起牛腹泻的病原体,在中国分布广泛,给养牛业造成了巨大损失。快速发现和准确鉴定BVDV对后续管理和流行病学调查至关重要。[方法]本研究以5'UTR基因为靶点,应用交叉引物扩增（CPA）与免疫层析试纸相结合的检测方法对BVDV进行检测。[结果]CPA检测限为每次反应640 fg,温度为62 ℃,反应时间为60 min。未观察到与牛感染的其他病原体发生交叉反应。此外,使用CPA测定法对100 份感染BVDV的现场样本进行准确评估。将CPA与常规PCR检测结果进行比较,结果显示,CPA比常规PCR具有更高的检出率。[结论]CPA检测是一种实用、有效的BVDV诊断工具。

关键词: 牛病毒性腹泻病毒（BVDV）, 交叉引物扩增（CPA）, 免疫层析, 诊断

Abstract: [Objective] Bovine viral diarrhea virus（BVDV）is a known causative agent of cattle diarrhea and is widely distributed across China. It caused huge losses to the cattle industry. Rapid detection and accurate identification of BVDV is essential for follow-up management and epidemiological investigations.[Method] In this study,targeting the 5'UTR gene,a cross-priming amplification combined with vertical flow （CPA-VF）assay was developed.[Result] The detection limit of the CPA assay was 640 fg per reaction at 62 ℃ for 60 min. No cross-reactions were observed with other pathogene infective to cattle. Furthermore,100 fecal sample from experimentally-infected animals were accurately assessed using the CPA assay. The performance of the CPA assay was compared with the results of conventional PCR.CPA had a higher detection rate than conventional PCR.[Conclusion] The CPA-VF assay is a practical and effective diagnostic tool for cattle diarrhea caused by BVDV.

Key words: bovine viral diarrhea virus（BVDV）, cross-priming amplification（CPA）, vertical flow assay, diagnose

[1] Srivastava M,Mittal L,Sarmadhikari D,et al.Template entrance channel as possible allosteric inhibition and resistance site for quinolines tricyclic derivatives in RNA dependent RNA polymerase of bovine viral diarrhea virus[J]. Pharmaceuticals (Basel),2023,16(3):376.
[2] Hong T Q,Yang L Y,Wang P Z,et al.Pestiviruses infection:Interferon-virus mutual regulation[J]. Front Cell Infect Microbiol,2023,13:1146394.
[3] Socha W, Larska M, Rola J,et al.Occurrence of bovine coronavirus and other major respiratory viruses in cattle in Poland[J]. Journal of Veterinary Research, 2022, 66(4):479-486.
[4] Rota P A,Oberste M S,Monroe S S,et al.Characterization of a novel coronavirus associated with severe acute respiratory syndrome[J]. Science,2003,300(5624):1394-1399.
[5] Drosten C, Muth D, Corman V M, et al. An observational,laboratory-based study of outbreaks of Middle East respiratory syndrome coronavirusin Jeddah and Riyadh,Kingdom of Saudi Arabia,2014[J]. Clinical Infectious Diseases,2015(3):369-377.
[6] Wang Q,Vlasova A N,Kenney S P,et al.Emerging and re-emerging coronaviruses in pigs[J]. Current Opinion Virology,2019,34:39-49.
[7] Furukawa S,Tsukamoto K,Maeda M.Multicentric histiocytosis related to avian leukosis virus subgroup J(ALV-J)-infection in meat-type local chickens[J].Journal of Veterinary Medical Science,2014,76,89-92.
[8] Brandao P E,Gregori F,Richtzenhain L J,et al.Molecular analysis of Brazilian strains of bovine coronavirus(BCoV) reveals a deletion within the hypervariable region of the S1 subunit of the spike glycoprotein also found in human coronavirus OC43[J]. Archives of Virology,2006,151:1735-1748.
[9] Yoo D W,Parker M D,Babiuk L A.The S2 subunit of the spike glycoprotein of bovine coronavirus mediates membrane fusion in insect cells[J].Virology,1991,180(1):395-399.
[10] Fehr A R,Perlman S.Coronaviruses:An overview of their replication and pathogenesis[J].Methods in Molecular Biology,2015,1282:1-23.
[11] Kubo H,Yamada Y K,Taguchi F.Localization of neutralizing epitopes and the receptor-binding site within the amino-terminal 330 amino acids of the murine coronavirus spike protein[J].Journal of Virology,1994,68(9):5403-5410.
[12] Cho K O,Hasoksuz M,Nielsen P R,et al.Cross-protection studies between respiratory and calf diarrhea and winter dysentery coronavirus strains in calves and RT-PCR and nested PCR for their detection[J].Archives of Virology,2001,146:2401-2419.
[13] Spencer J L,Chan M,Nadin-Davis S.Relationship between egg size and subgroup J avian leukosis virus in eggs from broiler breeders[J]. Avian Pathology Journal of the W.V.P.A,2000,29(6),617-622.
[14] Li D L,Qin L T,Gao H L,et al.Avian leukosis virus subgroup A and B infection in wild birds of Northeast China[J].Veterinary Microbiology,2013,163,257-263.
[15] Abraham G,Roeder P L,Zewdu R.Agents associated with neonatal diarrhoea in Ethiopian dairy calves[J].Tropical Animal Health and Production,1992,24(2),74-80.
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