Rapid on-the-spot detection of Edwardsiella piscicida using recombinase polymerase amplification with lateral flow

Jiatiao Jiang; Yuhang Fan; Sijie Zhang; Qiyao Wang; Yuanxing Zhang; Qin Liu; Shuai Shao

Aquaculture Reports (Feb 2022)

Rapid on-the-spot detection of Edwardsiella piscicida using recombinase polymerase amplification with lateral flow

Jiatiao Jiang,
Yuhang Fan,
Sijie Zhang,
Qiyao Wang,
Yuanxing Zhang,
Qin Liu,
Shuai Shao

Affiliations

Jiatiao Jiang: State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China
Yuhang Fan: State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China
Sijie Zhang: State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China
Qiyao Wang: State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China; Laboratory for Marine Biology and Biotechnology, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266071, China; Shanghai Engineering Research Center of Maricultured Animal Vaccines, 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, East China University of Science and Technology, Shanghai 200237, China; Laboratory for Marine Biology and Biotechnology, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266071, China; Shanghai Engineering Research Center of Maricultured Animal Vaccines, Shanghai 200237, China
Shuai Shao: State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China; Corresponding author.

Journal volume & issue: Vol. 22
p. 100945

Abstract

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Edwardsiella piscicida is one of the important pathogens in aquaculture, which takes huge losses to the economic benefits of turbot industry. Thus, a simple, rapid and accurate diagnostic assay of E. piscicida becomes an urgent need. In the present study, a detection method of E. piscicida based on recombinase polymerase amplification combined with lateral flow strip (RPA-LF) was established. The RPA-LF assay targeted E. piscicida evpP gene and could detect as low as 10 fg DNA of E. piscicida, which was approximately 1000 times more sensitive than conventional PCR assay. Meanwhile, the RPA-LF assay showed no cross-reactivity with DNA from other major bacterial pathogens in aquaculture. Besides, to improve the convenience of RPA-LF assay, a rapid on-the-spot sample preparation using nucleic acid extraction by cellulose dipsticks was established. Results indicated that bacteria could be detected as low as 1 copy and completed within 30 min. Moreover, this modified RPA-LF assay could be used to monitor E. piscicida infection in turbot industry.

Published in Aquaculture Reports

ISSN: 2352-5134 (Online)
Publisher: Elsevier
Country of publisher: Netherlands
LCC subjects: Agriculture: Aquaculture. Fisheries. Angling
Website: http://www.journals.elsevier.com/aquaculture-reports/

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