Dual recombinase polymerase amplification system combined with lateral flow immunoassay for simultaneous detection of Staphylococcus aureus and Vibrio parahaemolyticus
文献类型： 外文期刊
作者： Zhang, Yan 1 ; Liu, Xiaofeng 1 ; Luo, Jiawei 2 ; Liu, Hua 2 ; Li, You 2 ; Liu, Juan 5 ; Zhu, Lemei 5 ; Wang, Jinbin 2 ; Zeng, Haijuan 2 ;
作者机构： 1.Lanzhou Univ Technol, Sch Life Sci & Engn, Lanzhou 730050, Peoples R China
2.Shanghai Acad Agr Sci, Biotechnol Res Inst, Shanghai Profess Technol Serv Platform Agr Biosafe, Key Lab Agr Genet & Breeding, Shanghai 201106, Peoples R China
3.Minist Agr & Rural Affairs, Key Lab Safety Assessment Environm Agr Genet Modif, Shanghai 201106, Peoples R China
4.Shanghai Coelite Agr Scitech Grp Co Ltd, Shanghai 201106, Peoples R China
5.Changsha Med Univ, Sch Publ Hlth, Academician Workstat, Changsha, Peoples R China
关键词： Recombinase polymerase amplification; Lateral flow immunoassays; Foodborne pathogens; Multiplex detection
期刊名称：JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS （ 影响因子：3.1； 五年影响因子：3.1 ）
ISSN： 0731-7085
年卷期： 2025 年 255 卷
页码：
收录情况： SCI
摘要： Development of a highly sensitive visualization platform for multiplex genetic detection could significantly improve efficiency and reliability of on-site detection of foodborne pathogens. In this study, coupling recombinase polymerase amplification (RPA) with lateral flow immunoassay (LFIA) readout system was proposed for Staphylococcus aureus and Vibrio parahaemolyticus detection. Taking the advantage of the isothermal amplification of RPA, dual primers modified with different labeling groups were designed to realize target signal amplification. LFIA coated with anti-digoxigenin antibody and streptavidin as test line 1 and 2 were designed to detect the two RPA products. The proposed method (dual RPA-LFIA) could realize visual detection using LFIA through rapid RPA amplification within 20 min, exhibiting a lowest detection limit of 4.6 x 102 CFU/mL for Staphylococcus aureus and Vibrio parahaemolyticus. The dual RPA-LFIA is characterized by simultaneous detection of dual targets in one RPA reaction and colorimetric readout through LFIA, thus ensuring high sensitivity and efficiency, and showing great potential to address the on-site detection of foodborne pathogens in the future.
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