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Development of a real‐time recombinase polymerase amplification assay for rapid detection of Aeromonas hydrophila
Author(s) -
Qu Yang,
Wang Qing,
Li Yingying,
Wang Yingying,
Yin Jiyuan,
Ren Yan,
Liu Chun,
Liu Xiaofang,
Wang Yahui,
Zeng Weiwei
Publication year - 2021
Publication title -
journal of fish diseases
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.819
H-Index - 85
eISSN - 1365-2761
pISSN - 0140-7775
DOI - 10.1111/jfd.13291
Subject(s) - recombinase polymerase amplification , aeromonas hydrophila , biology , real time polymerase chain reaction , polymerase chain reaction , microbiology and biotechnology , taqman , aeromonas , virology , fish <actinopterygii> , gene , bacteria , fishery , genetics
Abstract Aeromonas hydrophila is ubiquitous in the aquaculture industry and a constant cause of severe disease and economic losses. The early diagnosis of these infections is crucial for disease surveillance and prevention. We developed a real‐time recombinase polymerase amplification (real‐time RPA) assay for detection of A. hydrophila using the haemolysin gene. The assay was performed at 37°C for 20 min and was highly specific with no cross‐reaction with other fish pathogens or with other Aeromonas species. The assay detection limit was 10 2 copies of the Aeromonas hydrophila per reaction. Compared with traditional culture‐based method or real‐time PCR, the diagnostic sensitivity and specificity of the real‐time RPA were 73.7 and 100%, as well as 64.7 and 93%. Our newly developed real‐time RPA was specific and sensitive and can be used in large‐scale and point‐of‐care field investigations of A. hydrophila infections to enable earlier diagnoses.