{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,9]],"date-time":"2026-05-09T00:42:52Z","timestamp":1778287372788,"version":"3.51.4"},"reference-count":35,"publisher":"Wiley","issue":"9","license":[{"start":{"date-parts":[[2022,1,7]],"date-time":"2022-01-07T00:00:00Z","timestamp":1641513600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["scijournals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["J Sci Food Agric"],"published-print":{"date-parts":[[2022,7]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>BACKGROUND<\/jats:title><jats:p><jats:italic>Klebsiella pneumoniae<\/jats:italic> is a zoonotic opportunistic pathogen, leading to severe infections in dairy cows and humans. Efficient, on\u2010site and accurate detection of <jats:italic>K. pneumoniae<\/jats:italic> is necessary to reduce the harm of cow mastitis and human infections. The objective of this study was to establish a recombinase\u2010aided amplification (RAA) method combined with lateral flow dipstick (LFD) for rapid detection of <jats:italic>K. pneumoniae<\/jats:italic>.<\/jats:p><\/jats:sec><jats:sec><jats:title>RESULTS<\/jats:title><jats:p>The primer concentration, incubation temperature and incubation time of the RAA reaction were optimized. When the primer concentration was 100\u2009nmol L<jats:sup>\u20131<\/jats:sup>, the strongest band could be obtained by incubation at 37\u2009\u00b0C for 20\u2009min. The RAA\u2010LFD method had high specificity to <jats:italic>K. pneumoniae<\/jats:italic> and showed no cross\u2010reaction with other pathogens. In addition, the detection limit of RAA\u2010LFD for <jats:italic>K. pneumoniae<\/jats:italic> was 20\u2009fg genomic DNA and 2.5\u2009\u00d7\u200910<jats:sup>2<\/jats:sup>\u2009CFU\u2009mL<jats:sup>\u22121<\/jats:sup> of bacteria in pure culture, which is 100 times higher than that of polymerase chain reaction (PCR) detection. Moreover, the RAA\u2010LFD method can detect <jats:italic>K. pneumoniae<\/jats:italic> at initial concentrations as low as 2.5\u00a0CFU per 25\u2009mL in artificially spiked milk samples after at least incubation for 6\u00a0h. Importantly, RAA\u2010LFD had a high agreement with a test accuracy of 96.9%, compared with the biochemical identification method. Also, the detection accuracy of RAA\u2010LFD was higher than that of the PCR assay (95.3%).<\/jats:p><\/jats:sec><jats:sec><jats:title>CONCLUSIONS<\/jats:title><jats:p>The results demonstrated that the RAA\u2010LFD assay is an accurate, sensitive, simple and point\u2010of\u2010use detection method for <jats:italic>K. pneumoniae<\/jats:italic>, which could be used as a potential application in the research laboratory and for disease diagnosis. \u00a9 2021 Society of Chemical Industry<\/jats:p><\/jats:sec>","DOI":"10.1002\/jsfa.11737","type":"journal-article","created":{"date-parts":[[2021,12,22]],"date-time":"2021-12-22T07:19:23Z","timestamp":1640157563000},"page":"3879-3886","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":21,"title":["Development of an isothermal recombinase\u2010aided amplification assay for the rapid and visualized detection of <i>Klebsiella 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