A recombinase polymerase amplification with lateral flow assay for rapid on-the-spot detection of Aeromonas salmonicida

Sijie Zhang; Yuanxing Zhang; Qin Liu; Qiyao Wang; Yibei Zhang

Water Biology and Security (Jul 2024)

A recombinase polymerase amplification with lateral flow assay for rapid on-the-spot detection of Aeromonas salmonicida

Sijie Zhang,
Yuanxing Zhang,
Qin Liu,
Qiyao Wang,
Yibei Zhang

Affiliations

Sijie Zhang: State Key Laboratory of Bioreactor Engineering, Laboratory for Aquatic Animal Diseases, East China University of Science and Technology, Shanghai, 200237, China
Yuanxing Zhang: Southern Marine Science and Engineering Guangdong Laboratory (Zhuhai), Zhuhai, 519000, China; Shanghai Engineering Research Center of Maricultured Animal Vaccines, Shanghai, 200237, China
Qin Liu: State Key Laboratory of Bioreactor Engineering, Laboratory for Aquatic Animal Diseases, East China University of Science and Technology, Shanghai, 200237, China; Southern Marine Science and Engineering Guangdong Laboratory (Zhuhai), Zhuhai, 519000, China; Shanghai Engineering Research Center of Maricultured Animal Vaccines, Shanghai, 200237, China
Qiyao Wang: State Key Laboratory of Bioreactor Engineering, Laboratory for Aquatic Animal Diseases, East China University of Science and Technology, Shanghai, 200237, China; Shanghai Engineering Research Center of Maricultured Animal Vaccines, Shanghai, 200237, China
Yibei Zhang: State Key Laboratory of Bioreactor Engineering, Laboratory for Aquatic Animal Diseases, East China University of Science and Technology, Shanghai, 200237, China; Shanghai Engineering Research Center of Maricultured Animal Vaccines, Shanghai, 200237, China; Corresponding author. State Key Laboratory of Bioreactor Engineering, Laboratory for Aquatic Animal Diseases, East China University of Science and Technology, Shanghai, 200237, China.

Journal volume & issue: Vol. 3, no. 3
p. 100272

Abstract

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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 °C. Its sensitivity extends to as low as 100 fg of purified DNA, representing more than 1000-fold 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/μL 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.

Published in Water Biology and Security

ISSN: 2772-7351 (Online)
Publisher: KeAi Communications Co. Ltd.
Country of publisher: China
LCC subjects: Geography. Anthropology. Recreation: Environmental sciences; Science: Biology (General)
Website: https://www.keaipublishing.com/en/journals/water-biology-and-security/

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