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Development of a Recombinase-aided Amplification Combined With Lateral Flow Dipstick Assay for the Rapid Detection of
Jiang Shuai Li1, Yan Zhe Hao2, Mei Ling Hou2
1Faculty of environment and life, Beijing University of Technology, Beijing 100024, China.
Objective:
To establish a sensitive, simple and rapid detection method for African swine fever virus (ASFV) B646L gene.
Methods:
A recombinase-aided amplification-lateral flow dipstick (RAA-LFD) assay was developed in this study. Recombinase-aided amplification (RAA) is used to amplify template DNA, and lateral flow dipstick (LFD) is used to interpret the results after the amplification is completed. The lower limits of detection and specificity of the RAA assay were verified using recombinant plasmid and pathogenic nucleic acid. In addition, 30 clinical samples were tested to evaluate the performance of the RAA assay.
Results:
The RAA-LFD assay was completed within 15 min at 37 °C, including 10 min for nucleic acid amplification and 5 minutes for LFD reading results. The detection limit of this assay was found to be 200 copies per reaction. And there was no cross-reactivity with other swine viruses.
Conclusion:
A highly sensitive, specific, and simple RAA-LFD method was developed for the rapid detection of the ASFV.
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