一种基于重组酶聚合酶扩增技术的琅琊病毒现场可视化检测方法

doi:10.1016/j.jia.2025.12.008

Journal of Integrative Agriculture ›› 2026, Vol. 25 ›› Issue (7): 3099-3101.DOI: 10.1016/j.jia.2025.12.008

一种基于重组酶聚合酶扩增技术的琅琊病毒现场可视化检测方法

收稿日期:2025-05-25 修回日期:2025-12-08 接受日期:2025-10-31 出版日期:2026-07-20 发布日期:2026-06-09

An on-site visual assay detecting Langya henipavirus based on recombinase polymerase amplification technology

Lin Cheng^{1, 2*}, Haili Zhang^3*, Xiaoai Zhang^4*, Pei Huang³, Xinlan Chen^{1, 2}, Fangxu Li¹, Wujian Li^{5, 6, 7}, Wei Liu^4#, Hualei Wang^3#, Sandra Chiu^{5, 6, 7#}, Zengguo Cao^1#

¹ State Key Laboratory of Virology and Biosafety, Wuhan Institute of Virology, Center for Biosafety Mega-Science, Chinese Academy of Sciences, Wuhan 430071, China
² University of Chinese Academy of Sciences, Beijing 100190, China
³ State 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
⁴ Beijing Institute of Microbiology and Epidemiology, Beijing 100101, China
⁵Department of Laboratory Medicine, The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei 230031, China
⁶ Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei 230026, China
⁷ Key Laboratory of Anhui Province for Emerging and Reemerging Infectious Diseases, Hefei 230026, China

Received:2025-05-25 Revised:2025-12-08 Accepted:2025-10-31 Online:2026-07-20 Published:2026-06-09
About author:#Correspondence Hualei Wang, E-mail: wanghualei@jlu.edu.cn; Wei Liu, E-mail: lwbime@163.com; Sandra Chiu, E-mail: qiux@ustc.edu.cn; Zengguo Cao, E-mail: caozg@wh.iov.cn * These authors contributed equally to this study.
Supported by:
This work was supported by the National Key Research and Development Program of China (2021YFF0703600).

摘要/Abstract

摘要：

琅琊病毒（Langya henipavirus, LayV）是一种新发的、人兽共患的亨尼帕病毒。副粘病毒科亨尼帕病毒属成员均具有极高的致病性，感染人和动物后可引起严重疾病，如同为亨尼帕病毒属的亨德拉病毒（Hendra virus, HeV）和尼帕病毒（Nipah virus, NiV）均可引起人类的高致死性疾病。为实现对LayV的高效、快速检测，本研究根据LayV核蛋白基因的保守序列，分别对基于重组酶聚合酶扩增原理设计的引物和探针进行特异性生物标记，并结合一次性核酸可视化检测装置，建立了一种针对LayV的现场可视化检测方法。将携带LayV核蛋白基因质粒梯度稀释后，作为评价方法的阳性模板，使用本研究建立的方法进行扩增，结果显示该方法的灵敏度可达1.22 copies/μL。使用HeV、NiV、仙台病毒（Sendai virus, SeV）的RNA作为扩增模板，评价该方法的特异性，结果显示，该方法对上述病毒无交叉反应，具有较好的特异性。此外，将在LayV感染的细胞和鼩鼱组织样品中提取的病毒RNA作为检测对象，分别使用建立的RT-qPCR方法和可视化检测方法进行检测，结果显示，两种方法的检测结果一致。但由于本研究建立的可视化检测方法无需特殊的仪器，且更易操作，因此更具实用性。同时开展双盲试验，使用本研究建立的可视化检测方法对LayV阳性和阴性的鼩鼱组织进行检测，并平行进行RT-qPCR方法检测，结果显示，可视化检测结果与RT-qPCR检测结果吻合的同时，与阳性和阴性样品的实际结果一致，证明本研究建立的可视化检测方法可以完成临床样品的检测。综上所述，上述检测方法具有操作简便、灵敏度高、特异性强等优势，作为检测LayV的可靠方法，具有较大的应用潜力，尤其适用于需要“现地快检”的应用场景。

Lin Cheng, Haili Zhang, Xiaoai Zhang, Pei Huang, Xinlan Chen, Fangxu Li, Wujian Li, Wei Liu, Hualei Wang, Sandra Chiu, Zengguo Cao. 一种基于重组酶聚合酶扩增技术的琅琊病毒现场可视化检测方法[J]. Journal of Integrative Agriculture, 2026, 25(7): 3099-3101.

Lin Cheng, Haili Zhang, Xiaoai Zhang, Pei Huang, Xinlan Chen, Fangxu Li, Wujian Li, Wei Liu, Hualei Wang, Sandra Chiu, Zengguo Cao. An on-site visual assay detecting Langya henipavirus based on recombinase polymerase amplification technology[J]. Journal of Integrative Agriculture, 2026, 25(7): 3099-3101.

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一种基于重组酶聚合酶扩增技术的琅琊病毒现场可视化检测方法

An on-site visual assay detecting Langya henipavirus based on recombinase polymerase amplification technology

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