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Title:Loop-mediated isothermal amplification : rapid molecular detection of virulence genes associated with avian pathogenic Escherichia coli in poultry
Authors:ID Kogovšek, Polona (Author)
ID Ambrožič, Jerneja (Author)
ID Dovč, Alenka (Author)
ID Dreo, Tanja (Author)
ID Hristov, Hristo (Author)
ID Krapež, Uroš (Author)
ID Ravnikar, Maja (Author)
ID Slavec, Brigita (Author)
ID Lotrič, Marjetka (Author)
ID Žel, Jana (Author)
ID Zorman-Rojs, Olga (Author)
Files:URL URL - Source URL, visit https://academic.oup.com/ps/advance-article/doi/10.3382/ps/pey516/5205916
 
.pdf PDF - Presentation file, download (177,60 KB)
MD5: 6F2A6B33A5AC5DC87C1FE69BA158F747
 
Language:English
Typology:1.01 - Original Scientific Article
Organization:Logo NIB - National Institute of Biology
Abstract: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.
Keywords:APEC virulence genes, isothermal amplification LAMP, on-site detection, colibacillosis
Publication status:Published
Publication version:Version of Record
Publication date:01.03.2019
Year of publishing:2018
Number of pages:str. 1500-1510
Numbering:Vol. 98, no. 3
PID:20.500.12556/DiRROS-19652 New window
UDC:577.2
ISSN on article:0032-5791
DOI:10.3382/ps/pey516 New window
COBISS.SI-ID:4878671 New window
Publication date in DiRROS:24.07.2024
Views:346
Downloads:151
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Record is a part of a journal

Title:Poultry science
Shortened title:Poultry sci.
Publisher:Oxford University Press
ISSN:0032-5791
COBISS.SI-ID:5679367 New window

Document is financed by a project

Funder:ARIS - Slovenian Research and Innovation Agency
Project number:P4-0165-2015
Name:Biotehnologija in sistemska biologija rastlin

Funder:ARIS - Slovenian Research and Innovation Agency
Project number:V4-1401-2014
Name:Obvladovanje okužb z E. coli pri perutnini: Določitev kritičnih mest vnosa bakterije E.coli, vključno z E. coli z ESBL v reje perutnine in študija preventivnih ukrepov za zmanjševanje porabe protimikrobnih zdravil

Funder:EC - European Commission
Project number:613908
Name:Development of Cost efficient Advanced DNA-based methods for specific Traceability issues and High Level On-site applicatioNs
Acronym:DECATHLON

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License:CC BY-NC-ND 4.0, Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International
Link:http://creativecommons.org/licenses/by-nc-nd/4.0/
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