Loop-mediated isothermal amplification : rapid molecular detection of virulence genes associated with avian pathogenic Escherichia coli in poultry

Kogovšek, Polona (author), Ambrožič, Jerneja (author), Dovč, Alenka (author), Dreo, Tanja (author), Hristov, Hristo (author), Krapež, Uroš (author), Ravnikar, Maja (author), Slavec, Brigita (author), Lotrič, Marjetka (author), Žel, Jana (author), Zorman-Rojs, Olga (author)

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.

• Language: English
• Keywords: APEC virulence genes, isothermal amplification LAMP, on-site detection, colibacillosis
• Work type: Article
• Typology: 1.01 - Original Scientific Article
• Organization: BF - Biotechnical Faculty; VF - Veterinary Faculty
• Publication status: Published
• Publication version: Version of Record
• Year: 2018
• Number of pages: Str. 1500-1510
• Numbering: Vol. 98, no. 3
• PID: 20.500.12556/RUL-144933
• UDC: 577.2
• ISSN on article: 0032-5791
• DOI: 10.3382/ps/pey516
• COBISS.SI-ID: 4878671
• Publication date in RUL: 23.03.2023

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

Licences

• License: CC BY 4.0, Creative Commons Attribution 4.0 International
• Link: http://creativecommons.org/licenses/by/4.0/
• Description: This is the standard Creative Commons license that gives others maximum freedom to do what they want with the work as long as they credit the author.
• Licensing start date: 23.03.2023