生物学杂志

生物学杂志 ›› 2025, Vol. 42 ›› Issue (6): 101-.doi: 10.3969/j.issn.2095-1736.2025.06.101

• 技术方法 • 上一篇    下一篇

甲型肝炎病毒RT-RPA结合侧流横向试纸条快检方法的建立

徐佳乐1， 汪艺涵2， 王永杰1,2,3， 喻勇新1,2   

  1. 1. 上海海洋大学 食品学院， 上海 201306；
    2. 农业农村部水产品质量安全贮藏保鲜风险评估实验室（上海）， 上海 201306；
    3. 青岛海洋科学与技术试点国家实验室 海洋生物学与生物技术功能实验室， 青岛 266237
  • 出版日期:2025-12-18 发布日期:2025-12-19
  • 通讯作者: 喻勇新，博士，高级工程师，研究方向为食源性致病微生物在环境及水产品中的传播机制及风险评估，E-mail:yxyu@shou.edu.cn
  • 作者简介:徐佳乐，硕士研究生，研究方向为食源性病毒，E-mail:2250119553@qq.com
  • 基金资助:
    上海市科委“科技创新行动计划”项目——水产品中食源性病毒快速多重RT-RPA-LFS检测产品的研发及应用（22N31900700）

Establishment of a rapid testing method for hepatitis A virus using RT-RPA combined with lateral flow strips

XU Jiale1, WANG Yihan2, WANG Yongjie1,2,3, YU Yongxin1,2   

  1. 1. College of Food Sciences and Technology, Shanghai Ocean University, Shanghai 201306, China; 2. Laboratory of
    Quality and Safety Risk Assessment for Aquatic Products on Storage and Preservation (Shanghai), Ministry of
    Agriculture and Rural Affairs, Shanghai 201306, China; 3. Marine Biology and Biotechnology Laboratory, Qingdao
    National Laboratory for Marine Science and Technology, Qingdao 266237, China
  • Online:2025-12-18 Published:2025-12-19

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摘要：
摘　要　研究针对甲型肝炎病毒（HAV）的快速检测需求，开发一种基于逆转录-重组酶聚合酶扩增（RT-RPA）技术结合侧流横向试纸条（LFS）的新型检测方法。从GenBank核苷酸数据库下载HAV Ⅰ~Ⅲ基因型序列，进行基因分型、分析，设计RT-RPA引物和探针。该方法能在42 ℃、20 min内完成HAV检测，灵敏度达到10 copies/μL，具有高度特异性，不与其他肠道病毒产生交叉反应，并且重复性达到71.4%，相较传统RT-qPCR技术，展现出更高的灵敏度和操作便捷性。RT-RPA-LFS检测技术的建立为HAV的现场快速诊断提供技术支持，对提高HAV检测效率、预防和控制HAV引起的食源性疫情具有重要意义。随着HAV疫苗的广泛使用，该检测技术的发展对区分疫苗株和野生株以及应对未来病毒多重检测的需求具有潜在的应用前景，为食品安全监测和公共卫生领域提供新的技术手段。

关键词: 甲型肝炎病毒, RT-RPA-LFS, 灵敏度, 特异性, 现场诊断

Abstract: A novel detection method for hepatitis A virus (HAV) was developed to meet the rapid detection needs, based on reverse transcription-recombinase polymerase amplification (RT-RPA) technology combined with lateral flow strips (LFS). Genotype Ⅰ-Ⅲ sequences of HAV were downloaded from the GenBank nucleotide database, and genetic typing and analysis were conducted to design primers and probes of RT-RPA. This method could complete HAV detection within 20 minutes at 42 ℃, with a sensitivity of 10 copies/μL, exhibiting high specificity, no cross-reactivity with other enteric viruses, and a repeatability of 71.4%. Compared to traditional RT-qPCR technology, this method demonstrated greater sensitivity and easier operation. The establishment of the RT-RPA-LFS detection technology provided technical support for on-site rapid diagnosis of HAV, which was of great significance for improving the efficiency of HAV detection and preventing and controlling foodborne outbreaks caused by HAV. With widespread HAV vaccine use, the development of this detection technology has potential applications for distinguishing vaccine strains from wild strains, as well as meeting the future needs for multiplex virus detection, providing new technical approaches for food safety monitoring and public health fields.

Key words: hepatitis A Virus, RT-RPA-LFS, sensitivity, specificity, on-site diagnosis

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