基于重组酶聚合酶扩增快速检测鸡传染性支气管炎病毒的胶体金试纸条

doi:10.11843/j.issn.0366-6964.2025.12.032

畜牧兽医学报 ›› 2025, Vol. 56 ›› Issue (12): 6299-6307.doi: 10.11843/j.issn.0366-6964.2025.12.032

基于重组酶聚合酶扩增快速检测鸡传染性支气管炎病毒的胶体金试纸条

郭鹏宇¹^,²^,³^,⁴(), 汤也¹^,²^,³^,⁴, 马丽华¹^,²^,³^,⁴, 孟宪臣¹^,²^,³^,⁴, 赵喆泓¹^,²^,³^,⁴, 谢泉¹^,²^,³^,⁴, 李拓凡¹^,²^,³^,⁴, 万志敏¹^,²^,³^,⁴, 邵红霞¹^,²^,³, 秦爱建¹^,²^,³, 叶建强¹^,²^,³^,⁴^,*()

1. 扬州大学兽医学院, 禽类预防医学教育部重点实验室, 江苏省动物预防医学重点实验室, 扬州 225009
2. 江苏高校动物重要疫病与人兽共患病防控协同创新中心, 扬州 225009
3. 扬州大学农业科技发展研究院(国际联合实验室), 扬州 225009
4. 教育部农业与农产品安全国际合作联合实验室, 扬州 225009

收稿日期:2025-02-18 出版日期:2025-12-23 发布日期:2025-12-24
通讯作者: 叶建强 E-mail:850189125@qq.com;jqye@yzu.edu.cn
作者简介:郭鹏宇(1998-)，女，黑龙江哈尔滨人，博士，主要从事禽传染性支气管炎相关研究，E-mail：850189125@qq.com
基金资助:
江苏高校优势学科建设工程资助项目

Colloidal Gold Test Strip for Rapid Detection of Chicken Infectious Bronchitis Virus based on Recombinase Polymerase Amplification

GUO Pengyu¹^,²^,³^,⁴(), TANG Ye¹^,²^,³^,⁴, MA Lihua¹^,²^,³^,⁴, MENG Xianchen¹^,²^,³^,⁴, ZHAO Zhehong¹^,²^,³^,⁴, XIE Quan¹^,²^,³^,⁴, LI Tuofan¹^,²^,³^,⁴, WAN Zhimin¹^,²^,³^,⁴, SHAO Hongxia¹^,²^,³, QIN Aijian¹^,²^,³, YE Jianqiang¹^,²^,³^,⁴^,*()

1. Key Laboratory of Jiangsu Preventive Veterinary Medicine, Key Laboratory for Avian Preventive Medicine, Ministry of Education, College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China
2. Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou 225009, China
3. Institute of Agricultural Science and Technology Development, Yangzhou University, Yangzhou 225009, China
4. Joint International Research Laboratory of Agriculture and Agri-Product Safety, the Ministry of Education of China, Yangzhou University, Yangzhou 225009, China

Received:2025-02-18 Online:2025-12-23 Published:2025-12-24
Contact: YE Jianqiang E-mail:850189125@qq.com;jqye@yzu.edu.cn

摘要/Abstract

摘要：

鸡传染性支气管炎是由鸡传染性支气管炎病毒(IBV)感染引起的鸡的一种急性、高度传染性的病毒性呼吸道疾病。为建立一种适合现场化检测IBV的方法，本研究靶向IBV的保守基因M设计引物及探针，结合RPA与胶体金技术，创制了一种能在1 h内出结果的检测IBV的RPA-胶体金试纸条方法。特异性检测证明，该RPA-试纸条方法仅能检测出IBV，而不与禽流感病毒(AIV)、鸡传染性法氏囊病毒(IBDV)、新城疫病毒(NDV)、禽白血病病毒(ALV)和鹅星状病毒(GAstV)等反应。灵敏度检测证明，该RPA-试纸条方法的最低检测限为8.26 TCID₅₀·mL^-1。对31份临床样本的比对检测表明，本RPA-试纸条方法、qPCR以及常规PCR的检测阳性率分别为74.2%、64.5%与54.8%。综上，本研究建立的RPA-试纸条快速检测IBV的方法特异性强、敏感性高、操作便捷，为IBV高效诊断及其免疫防控提供了新方法。

关键词: 鸡传染性支气管炎病毒, M基因, RPA, 胶体金试纸条

Abstract:

Infectious bronchitis is an acute and highly contagious respiratory disease caused by infectious bronchitis virus (IBV). In order to establish a suitable method for the field detection of IBV, we designed primers and probes targeting M, a conserved gene of IBV, and combined RPA and colloidal gold technology to create an RPA-test strip method for IBV detection within 1 hour. The specificity test demonstrated that the RPA-test strip method only detected IBV, but did not react with the avian influenza virus (AIV), chicken bursal disease virus (IBDV), Newcastle disease virus (NDV), avian leukosis virus (ALV), and goose astrovirus (GAstV). The sensitivity test proved that the lowest detection line of this RPA-test strip method was 8.26 TCID₅₀·mL^-1. Comparison of 31 clinical samples showed that the positive detection rates of this RPA-strip method, qPCR and conventional PCR were 74.2%, 64.5% and 54.8%, respectively. The above data showed that the RPA-strip method for the rapid detection of IBV established in this study is highly specific, sensitive and easy to operate, which provides a new method and technology for the efficient diagnosis of IBV and its immunological prevention and control.

Key words: infectious bronchitis virus, M gene, RPA, colloidal gold immunochromatographic test strip

中图分类号:

S858.315.3

郭鹏宇, 汤也, 马丽华, 孟宪臣, 赵喆泓, 谢泉, 李拓凡, 万志敏, 邵红霞, 秦爱建, 叶建强. 基于重组酶聚合酶扩增快速检测鸡传染性支气管炎病毒的胶体金试纸条[J]. 畜牧兽医学报, 2025, 56(12): 6299-6307.

GUO Pengyu, TANG Ye, MA Lihua, MENG Xianchen, ZHAO Zhehong, XIE Quan, LI Tuofan, WAN Zhimin, SHAO Hongxia, QIN Aijian, YE Jianqiang. Colloidal Gold Test Strip for Rapid Detection of Chicken Infectious Bronchitis Virus based on Recombinase Polymerase Amplification[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(12): 6299-6307.

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引物 Primers	序列(5′→3′) Sequences(5′→3′)	位置 Gene location
IBV-F	CGATAATACTTACTGTGTTTGCGTG	245—269
IBV-R	[biotin]-AAAGACTACTTCCTCCTGTTGCTAC	646—670
IBV probe	[FAM]-GAGCAATGCCGTAGGTTCAATACTCCTAACA [THE]AATGGTCAACAATGTA-[C3spacer]	348—394

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基于重组酶聚合酶扩增快速检测鸡传染性支气管炎病毒的胶体金试纸条

Colloidal Gold Test Strip for Rapid Detection of Chicken Infectious Bronchitis Virus based on Recombinase Polymerase Amplification

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项目 Item		常规PCR		总数 Total number	qPCR		总数 Total number
项目 Item		阳性Positive	阴性Negative	总数 Total number	阳性Positive	阴性Negative	总数 Total number
RPA	阳性Positive	13	10	23	18	5	23
	阴性Negative	4	4	8	2	6	8
总数Total number		17	14	31	20	11	31