| 165 | 0 | 16 |
| 下载次数 | 被引频次 | 阅读次数 |
为构建水貂阿留申病毒(Aleutian mink disease virus,AMDV)的快速、精准、定量检测方法，根据VP2蛋白的保守区域设计特异性引物和探针，构建了TaqMan qPCR检测方法。结果表明：该方法可对AMDV进行特异性、高敏感性及可重复性定量检测。质粒DNA最低检测限为3.45×101 copies/μL，敏感性是常规PCR的100倍；批次内和批次间变异系数均<2.00%。对来自山东省不同地区的水貂临床样本进行TaqMan qPCR检测，结果显示，总阳性率为76%,AMDV感染率较高。采用构建的方法对水貂不同组织进行检测，结果显示，AMDV在各组织中均有分布，脾脏和淋巴结中病毒载量较高(6.84×108 copies/g,4.99×108 copies/g)，脑组织中病毒载量较低(1.99×107 copies/g)。说明本研究构建的TaqMan qPCR检测方法可用于AMDV的快速、定量检测，研究结果为进一步揭示AMDV的组织嗜性和致病机制提供理论依据。
Abstract：To establish a rapid, precise, and quantitative detection method for Aleutian mink disease virus(AMDV), specific primers and probes were designed based on the conserved region of the VP2 protein, and a TaqMan qPCR detection method was established, achieving specific, highly sensitive, and repeatable detection and quantification of AMDV. The detection limit of plasmid DNA was as low as 3.45×101 copies/μL, and the sensitivity was 100 times than that of conventional PCR. The intra-batch and inter-batch coefficient of variation were both less than 2.00%. TaqMan qPCR was used to detect clinical samples from minks in different regions of Shandong Province. The results showed that the total positive rate in Shandong Province was 76%, and the prevalence of AMDV infection was relatively high. The AMDV TaqMan qPCR method was used to detect different tissues of minks. The results showed that AMDV was distributed in all organs, with higher viral loads in the spleen and lymph nodes(6.84×108 copies/g, 4.99×108 copies/g), and a lower viral load in the brain tissue(1.99×107 copies/g). The TaqMan qPCR detection method established in this study could be used for rapid and quantitative detection of AMDV. The results provide a theoretical basis for further revealing the tissue tropism and pathogenic mechanism of AMDV.
[1]赵永强，吴艳虹，章秀婷，等.貉源阿留申病毒全基因组测序及末端序列分子特征分析[J].病毒学报，2021, 37(2):389-397.
[2]Clemens D L, Wolfinbarger J B, Mori S, et al. Expression of Aleutian mink disease parvovirus capsid proteins by a recombinant vaccinia virus:Self-assembly of capsid proteins into particles[J]. Journal of Virology, 1992, 66(5):3077-3085.
[3]Vahedi S M, Salek Ardestani S, Banabazi M H, et al. Epidemiology, pathogenesis, and diagnosis of Aleutian disease caused by Aleutian mink disease virus:A literature review with a perspective of genomic breeding for disease control in American mink(Neogale Vison)[J]. Virus Research, 2023, 336:199208.
[4]吴占运.水貂阿留申病的危害及防控措施[J].中国动物保健，2022, 24(1):79-80.
[5]Tong M W, Sun N, Cao Z G, et al. Molecular epidemiology of Aleutian mink disease virus from fecal swab of mink in Northeast China[J]. BMC Microbiology, 2020, 20(1):234.
[6]于华玲，崔卓荣.水貂阿留申病的诊断与防控[J].山东畜牧兽医，2023, 44(7):38-39.
[7]卞大伟.水貂阿留申细小病毒的致病机制、鉴定及防治方法[J].山东畜牧兽医，2017, 38(4):62-65.
[8]Jepsen J R, D′Amore F, Baandrup U, et al. Aleutian mink disease virus and humans[J]. Emerging Infectious Diseases,2009, 15(12):2040-2042.
[9]韦韬，吴艳虹，丛丽，等.水貂阿留申病诊断技术研究进展[J].动物医学进展，2019, 40(6):78-82.
[10]廉慧锋，胡传伟，李叶，等.一株水貂阿留申病细小病毒的分离与鉴定[J].山东畜牧兽医，2013, 34(11):8-9, 10.
[11]Li L, Hu Z, Sun J H, et al. Development of an Eva Greenbased real-time PCR assay for detection of Aleutian mink disease virus[J]. Journal of Virological Methods, 2020, 275:113751.
[12]Cho H J, Ingram D G. Antigen and antibody in aleutian disease in mink. I. precipitation reaction by agar-gel electrophoresis[J]. The Journal of Immunology, 1972, 108(2):555-557.
[13]Knuuttila A, Aronen P, Saarinen A, et al. Development and evaluation of an enzyme-linked immunosorbent assay based on recombinant VP2 capsids for the detection of antibodies to Aleutian mink disease virus[J]. Clinical and Vaccine Immunology, 2009, 16(9):1360-1365.
[14]张秀丽，于慧娟，徐超，等.水貂阿留申病毒荧光定量PCR检测方法的建立及初步应用[J].中国畜牧兽医，2014, 41(9):1-5.
[15]许裕敬，卢雨萌，刘祎璇，等.水貂源大肠杆菌磺胺类耐药基因的检测与分析[J].经济动物学报，2023, 27(4):251-254.
[16]Cao Z, Xu H, Zhao X R, et al. Multiplex one-step RT-qPCR assays for simultaneous detection of AMDV, MEV and CDV[J]. BMC Veterinary Research, 2025, 21(1):18.
[17]王春霞，商金源，吴顺，等.水貂阿留申病毒微滴式数字PCR检测方法的建立与应用[J].中国预防兽医学报，2024, 46(9):931-937.
[18]李俚，胡哲，孙金辉，等.水貂阿留申病诊断技术的进展[J].中国兽医杂志，2018, 54(11):58-60.
[19]汪婷婷，马青霞，刘宏莹，等.水貂阿留申病病毒研究进展[J].中国畜牧兽医，2023, 50(8):3354-3363.
[20]吴艳虹，韦韬，丛丽，等.肉食兽阿留申病毒VP2和NS1蛋白功能区回顾及比较分析[J].病毒学报，2020, 36(4):726-734.
[21]张定秀，井长华，王若楠，等.山东省部分地区水貂阿留申病PCR检测及病原的分离鉴定[J].经济动物学报，2025, 29(4):299-304.
[22]孟庆峰，梁艳婷，肖成蕊，等.水貂阿留申病毒部分VP2基因的克隆表达和重组蛋白的免疫学分析[J].吉林农业大学学报，2009, 31(3):330-333.
[23]Yang Z X, Li Y F, Jiang Y X, et al. A developed TaqMan probe-based qPCR was used to quantify the distribution of AMDV in various tissues of infected mink and its prevalence in Northern China[J]. Frontiers in Veterinary Science, 2025,11:1498481.
基本信息:
DOI：10.13326/j.jea.2026.2228
中图分类号:S852.65
引用信息:
[1]戚少华,赵鑫,聂建鸿,等.水貂阿留申病毒TaqMan qPCR检测方法的建立及其在水貂组织中的分布[J].经济动物学报,2026,30(02):144-149.DOI:10.13326/j.jea.2026.2228.
基金信息:
山东省现代农业产业技术体系建设专项（SDARS-21-13）; 农业农村部动物疫病东营野外科学观测研究站项目（MARAORS260-AH023）
2026-01-08
2026-01-08
2026-01-08