20.500.12556/RUL-144933 Loop-mediated isothermal amplification rapid molecular detection of virulence genes associated with avian pathogenic Escherichia coli in poultry Infections with pathogenic Escherichia colican lead to different animal- and human-associated diseases. E. coliinfections are common in intensive poultry farming, and important economic losses can be expected during infections with avian pathogenic E. coli(APEC) strains followed by colibacillosis. Loop-mediated isothermal amplification (LAMP) assays were developed for rapid detection of 3 APEC-associated virulence genes: sitA, traT, and ompT. All 3 LAMP assays are shown to be specific, repeatable, and reproducible. High sensitivities of the assays are shown, where as few as 1,000 bacterial cells/mL can be detected in different matrices. On-site applicability of this LAMP method is demonstrated through testing of different sample types, from animal swabs and tissues, and from environmental samples collected from 6 commercial poultry farms. All 3 virulence genes were detected at high rates (above 85%) in samples from layer and broiler chickens with clinical signs and, interestingly, high prevalence of those genes was detected also in samples collected from clinically healthy broiler flock (above 75%) while lower prevalence was observed in remaining 3 clinically healthy chicken flocks (less than 75%). Importantly, these virulence genes were detected in almost all of the air samples from 11 randomly selected poultry houses, indicating air as an important route of E. colispread. Three LAMP assays that target APEC-associated virulence genes are shown to be sensitive and robust and are therefore applicable for rapid on-site testing of various sample types, from animal swabs to air. This on-site LAMP testing protocol offers rapid diagnostics, with results obtained in <35 min, and it can be applied to other important microorganisms to allow the required prompt measures to be taken. APEC virulence genes isothermal amplification LAMP on-site detection colibacillosis true false true Angleški jezik Ni določen Članek v reviji 2023-03-23 13:04:38 2023-03-23 13:04:39 2023-03-24 03:37:40 0000-00-00 00:00:00 2018 0 0 Str. 1500-1510 no. 3 Vol. 98 2019 0000-00-00 Zaloznikova Objavljeno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 577.2 0032-5791 10.3382/ps/pey516 4878671 Loop-mediated-Zorman_Rojs.pdf Loop-mediated-Zorman_Rojs.pdf 1 763F8A68CAEF818CEB7EAB6B6B4FB0AD 7984502871742cd4deeee4a2d5099288a36775cb305bbbadba9a41898102e7c6 29401c60-c974-11ed-a314-00155dcfd717 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=167506 https://academic.oup.com/ps/advance-article/doi/10.3382/ps/pey516/5205916 1 cf190225-c972-11ed-a314-00155dcfd717 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=167505 Biotehniška fakulteta Veterinarska fakulteta 0 0 0