尼帕病毒RT-RAA-VF核酸可视化检测方法的建立

doi:10.1016/j.jia.2024.11.018

Journal of Integrative Agriculture ›› 2025, Vol. 24 ›› Issue (2): 790-794.DOI: 10.1016/j.jia.2024.11.018

尼帕病毒RT-RAA-VF核酸可视化检测方法的建立

收稿日期:2024-04-19 接受日期:2024-09-24 出版日期:2025-02-20 发布日期:2025-01-22

On-site visual detection of Nipah virus combining a reverse transcription recombinase-aided amplification with a lateral-flow dipstick assay

Kaikai Jin^1*, Junjie Zhao^{1, 3*}, Huanxin Chen^1*, Zimo Zhang¹, Zengguo Cao², Zanheng Huang¹, Hao Li¹, Yongsai Liu¹, Lisi Ai¹, Yufei Liu¹, Changqi Fan⁴, Yuanyuan Li¹, Pei Huang^1#, Hualei Wang^1#, Haili Zhang^1#

¹State Key Laboratory for Diagnosis and Treatment of Severe Zoonotic Infectious Diseases/Key Laboratory for Zoonosis Research of the Ministry of Education/Institute of Zoonosis, College of Veterinary Medicine, Jilin University, Changchun 130062, China
²Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Center for Biosafety Mega-Science, Chinese Academy of Sciences, Wuhan 430071, China
³National/WOAH Reference Laboratory for Classical Swine Fever, China Institute of Veterinary Drug Control, Beijing 100081, China
⁴Zhili College, Tsinghua University, Beijing 100084, China

Received:2024-04-19 Accepted:2024-09-24 Online:2025-02-20 Published:2025-01-22
About author:Kaikai Jin, E-mail: 1312467960@qq.com; Junjie Zhao, E-mail: 1229195209@qq.com; Huanxin Chen, E-mail: 1852384295@qq.com; #Correspondence Hualei Wang, E-mail: wanghualei@jlu.edu.cn; Pei Huang, E-mail: huangpei@jlu.edu.cn; Haili Zhang, E-mail: zhanghaili@jlu.edu.cn * These authors contributed equally to this work.
Supported by:
This study was supported by the National Key Research and Development Program of China (2021YFF0703600).

摘要/Abstract

摘要：

尼帕病毒（NiV）是一种新兴的人畜共患病毒，具有广泛的宿主范围，以果蝠为主要自然宿主，可感染人类及猪、马、狗等多种动物。NiV可通过多种途径感染人类和其他动物，包括接触被感染的动物及其体液，食用被污染的食物或水源等途径传播，且存在人与人之间传播的可能性。NiV感染的人类和动物会出现严重的呼吸系统和中枢神经系统症，尤为严重的是，NiV感染人类的致死率高达40-75%，这也使其成为全球公共卫生领域的一大挑战。鉴于目前尚没有针对NiV感染人类或动物的有效治疗手段和疫苗，研发一种能够实现NiV早期检测的方法显得尤为迫切，这对于控制疫情的爆发和传播至关重要。在本研究中，我们比对了55个不同NiV毒株的P基因序列，选择了 P 基因的高度保守区作为靶标，设计了特异性的重组酶介导等温核酸扩增技术（RAA）引物和探针。结合侧向免疫层析试纸条（VF），建立了一种快速、特异、灵敏的NiV核酸可视化检测方法（RT-RAA-VF）。该方法有效减少了气溶胶污染的风险，并能够在42 ℃下，在20分钟检测到低至5 copies μL^-1的尼帕病毒RNA转录本，具有较高的灵敏度。该检测方法对NiV具有高度特异性，与亨德拉病毒（HeV）、狂犬病病毒（RABV）、单纯疱疹病毒1型（HSV-1）以及猪链球菌等其他可引起NiV感染类似临床症状的病原体无交叉反应。此外，在模拟临床样本的检测中，本方法与世界动物卫生组织（WOAH）推荐的实时RT-PCR方法显示出100 % 的符合率。以上结果表明，本研究建立的针对NiV P 基因的RT-RAA-VF检测方法有望成为NiV感染早期和现场诊断的有效工具。

Kaikai Jin, Junjie Zhao, Huanxin Chen, Zimo Zhang, Zengguo Cao, Zanheng Huang, Hao Li, Yongsai Liu, Lisi Ai, Yufei Liu, Changqi Fan, Yuanyuan Li, Pei Huang, Hualei Wang, Haili Zhang. 尼帕病毒RT-RAA-VF核酸可视化检测方法的建立[J]. Journal of Integrative Agriculture, 2025, 24(2): 790-794.

Kaikai Jin, Junjie Zhao, Huanxin Chen, Zimo Zhang, Zengguo Cao, Zanheng Huang, Hao Li, Yongsai Liu, Lisi Ai, Yufei Liu, Changqi Fan, Yuanyuan Li, Pei Huang, Hualei Wang, Haili Zhang. On-site visual detection of Nipah virus combining a reverse transcription recombinase-aided amplification with a lateral-flow dipstick assay[J]. Journal of Integrative Agriculture, 2025, 24(2): 790-794.

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尼帕病毒RT-RAA-VF核酸可视化检测方法的建立

On-site visual detection of Nipah virus combining a reverse transcription recombinase-aided amplification with a lateral-flow dipstick assay

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