A 型塞内卡病毒抗体检测胶体金免疫层析试纸条的制备及初步应用

doi:10.11843/j.issn.0366-6964.2025.08.045

畜牧兽医学报 ›› 2025, Vol. 56 ›› Issue (8): 4086-4094.doi: 10.11843/j.issn.0366-6964.2025.08.045

A 型塞内卡病毒抗体检测胶体金免疫层析试纸条的制备及初步应用

李帅鹏¹^,²(), 石正旺², 陈婕², 罗俊聪², 朱昱茜¹^,², 石鑫泰², 席韬², 张帆², 何印娣¹^,², 郑海学¹^,²^,*(), 张小丽¹^,*(), 田宏²^,*()

1. 甘肃农业大学动物医学院, 兰州 730070
2. 中国农业科学院兰州兽医研究所兰州大学动物医学与生物安全学院动物疫病防控全国重点实验室, 兰州 730000

收稿日期:2024-09-10 出版日期:2025-08-23 发布日期:2025-08-28
通讯作者: 郑海学,张小丽,田宏 E-mail:2962386234@qq.com;zhenghaixue@caas.cn;zhxl228@qq.com;xibeitian0931@163.com
作者简介:李帅鹏(2000-)，男，河南许昌人，硕士，主要从事动物疫病诊断技术研究，E-mail：2962386234@qq.com
基金资助:
中央高校基本科研经费专项资金(1ZUJBKY-2022-CT02)；国家生猪技术创新中心(NCTIP-XD/C03)；The STI 2030-Major Projects (2023ZD0404301)；2023农业技术产业体系建设(CARS-35)；中国农业科学院创新计划(CAAS-CSLPDCP-202302)；甘肃省重大科技专项项目(23ZDNA007)

Preparation and Preliminary Application of Colloidal Gold Immunochromatographic Test Strip for Senecavirus A Antibody Detection

LI Shuaipeng¹^,²(), SHI Zhengwang², CHEN Jie², LUO Juncong², ZHU Yuqian¹^,², SHI Xintai², XI Tao², ZHANG Fan², HE Yindi¹^,², ZHENG Haixue¹^,²^,*(), ZHANG Xiaoli¹^,*(), TIAN Hong²^,*()

1. College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, China
2. State Key Laboratory of Animal Disease Prevention and Control, College of Animal Medicine and Biosecurity, Lanzhou University, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730000, China

Received:2024-09-10 Online:2025-08-23 Published:2025-08-28
Contact: ZHENG Haixue, ZHANG Xiaoli, TIAN Hong E-mail:2962386234@qq.com;zhenghaixue@caas.cn;zhxl228@qq.com;xibeitian0931@163.com

摘要/Abstract

摘要：

本研究拟建立一种快速、准确、简便的A 型塞内卡病毒(Senecavirus A, SVA)抗体胶体金检测方法，用于SVA的感染诊断。通过原核系统表达纯化获得SVA VP2重组蛋白，并制备VP2蛋白多克隆抗体；将VP2蛋白与胶体金偶联形成金标抗原。随后，将VP2蛋白和VP2多克隆抗体分别包被在硝酸纤维素膜(NC膜)上作为检测线(T线)和质控线(C线)，优化反应体系制备试纸条。结果表明，制备的试纸条检测口蹄疫病毒O型、口蹄疫病毒A 型、猪繁殖与呼吸综合征病毒、猪瘟病毒、猪伪狂犬病病毒、非洲猪瘟病毒阳性血清及健康猪(SPF)血清时，均无交叉反应，特异性良好；对SVA阳性血清的检测灵敏度达1 ∶64，通过对150份猪临床血清样品进行试纸条和病毒中和试验的平行检测，两者的Kappa值为0.88，表明两种方法高度一致。综上所述，本研究成功开发了SVA抗体检测试纸条，可在10~15 min内完成检测，具有快速、准确、简便等优点，为临床定性检测及现场快速诊断塞内卡病毒病提供了有效工具。

关键词: 塞内卡病毒抗体, VP2蛋白, 胶体金免疫层析试纸条

Abstract:

The aim of this study was to develop a rapid, accurate and easy-to-use colloidal gold assay for Senecavirus A (SVA) antibody diagnosis for Senecavirus infection. The recombinant protein of SVA VP2 was obtained by prokaryotic expression and purification, and the VP2 protein polyclonal antibody was prepared. The VP2 protein was coupled with colloidal gold to form a gold labeled antigen. Subsequently, VP2 protein and VP2 polyclonal antibody were coated on nitrocellulosic membrane (NC membrane) as test line (T-line) and control line (C-line), respectively. Optimizing the reaction system to prepare test strips. The results showed that the prepared test strips had no cross reactivity and good specificity when detecting foot-and-mouth disease virus (FMDV) serotypes O and A (FMDV-O, FMDV-A), porcine reproductive and respiratory syndrome virus (PRRSV), classical swine fever virus (CSFV), pseudorabies virus (PRV), African swine fever virus (ASFV) positive serum and healthy pig (SPF) serum; The sensitivity of SVA positive serum was 1 ∶64. The kappa value of the strip and virus neutralization test (VNT) was 0.88 in 150 swine clinical samples, indicating that the two methods were highly consistent. In summary, an SVA antibody test strip was successfully developed by this study, which can be operated in 10 to 15 minutes, and has the advantages of being rapid, accurate and simple, providing an effective tool for clinical qualitative detection and on-site diagnosis of Senecavirus disease.

Key words: Senecavirus antibodies, VP2 protein, colloidal gold immunochromatographic test strip

中图分类号:

S852.659.6

李帅鹏, 石正旺, 陈婕, 罗俊聪, 朱昱茜, 石鑫泰, 席韬, 张帆, 何印娣, 郑海学, 张小丽, 田宏. A 型塞内卡病毒抗体检测胶体金免疫层析试纸条的制备及初步应用[J]. 畜牧兽医学报, 2025, 56(8): 4086-4094.

LI Shuaipeng, SHI Zhengwang, CHEN Jie, LUO Juncong, ZHU Yuqian, SHI Xintai, XI Tao, ZHANG Fan, HE Yindi, ZHENG Haixue, ZHANG Xiaoli, TIAN Hong. Preparation and Preliminary Application of Colloidal Gold Immunochromatographic Test Strip for Senecavirus A Antibody Detection[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(8): 4086-4094.

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参考文献 36

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方法 Method		病毒中和试验 Virus neutralization test
方法 Method		阳性 Positive	阴性 Negative	合计 Total
试纸条 Test strip	阳性 Positive	13	3	16
	阴性 Negative	0	134	134
	合计 Total	13	137	150
Kappa值 Kappa value	0.88

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A 型塞内卡病毒抗体检测胶体金免疫层析试纸条的制备及初步应用

Preparation and Preliminary Application of Colloidal Gold Immunochromatographic Test Strip for Senecavirus A Antibody Detection

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