文献类型：期刊文献

英文题名：A recombinase polymerase ampli fi cation with lateral fl ow assay for rapid on-the-spot detection of Aeromonas salmonicida

作者：Zhang, Sijie[1];Zhang, Yuanxing[2,3];Liu, Qin[1,2,3];Wang, Qiyao[1,3];Zhang, Yibei[1,3]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, Lab Aquat Anim Dis, Shanghai 200237, Peoples R China;[2]Southern Marine Sci & Engn Guangdong Lab Zhuhai, Zhuhai 519000, Peoples R China;[3]Shanghai Engn Res Ctr Maricultured Anim Vaccines, Shanghai 200237, Peoples R China

年份：2024

卷号：3

期号：3

外文期刊名：WATER BIOLOGY AND SECURITY

收录：WOS:【ESCI(收录号:WOS:001273451900001)】；

基金：Funding This work was fi nancially supported by National Key Research and Development Program of China (2022YFE0101200) , National Natural Science Foundation of China (32102850) , and Shanghai Agricultural Science and Technology Innovation Project (T2023328) .

语种：英文

外文关键词：Aeromonas salmonicida; Recombinase Polymerase Amplification (RPA); Lateral flow; On-the-spot detection

摘要：Aeromonas salmonicida is a common pathogen of salmonid fishes that poses a significant threat to the fresh water and marine culture industry, potentially resulting in huge economic losses. To prevent and control fish diseases caused by A. salmonicida , rapid and effective diagnostic approaches must be developed, and which are important for routine monitoring and clinical care. By combining recombinase polymerase amplification (RPA) technology with a visible lateral flow strip (RPA-LF), we have enhanced both the precision of RPA detection and the convenience of real-time monitoring. In this study, we introduce a robust method for detecting A. salmonicida using RPA-LF. This assay specifically targets the ASA_1441 gene of A. salmonicida , ensuring high specificity, without cross-reactivity with other prevalent fresh water or marine pathogens. The optimal amplification temperature of the RPA assay was 39 degrees C. Its sensitivity extends to as low as 100 fg of purified DNA, representing more than 1000fold higher sensitivity than conventional PCR methods. Furthermore, to enhance the usability of the RPA-LF assay, we developed a rapid sample preparation method using cellulose dipsticks for nucleic acid extraction. This method achieves a limit of detection (LOD) as low as 1.67 CFU/ mu L and completes the entire process within 20 min. In conclusion, our findings present a rapid and precise tool for monitoring A. salmonicida infection in aquaculture and marine culture. This advancement offers valuable insights for effective disease prevention and control strategies.