Development of a multiplex-PCR for rapid detection of the enteric pathogens Lawsonia intracellularis, Brachyspira hyodysenteriae, and Brachyspira pilosicoli in porcine faeces

T. La; A.M. Collins; N.D. Phillips; A. Oksa; D.J. Hampson

doi:10.1111/j.1472-765X.2005.01831.x

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Development of a multiplex-PCR for rapid detection of the enteric pathogens Lawsonia intracellularis, Brachyspira hyodysenteriae, and Brachyspira pilosicoli in porcine faeces

Journal article

Peer reviewed

Development of a multiplex-PCR for rapid detection of the enteric pathogens Lawsonia intracellularis, Brachyspira hyodysenteriae, and Brachyspira pilosicoli in porcine faeces

T. La, A.M. Collins, N.D. Phillips, A. Oksa and D.J. Hampson

Letters in Applied Microbiology, Vol.42(3), pp.284-288

2006

DOI: https://doi.org/10.1111/j.1472-765X.2005.01831.x

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Abstract

Aims: To develop an assay to simultaneously detect Lawsonia intracellularis, Brachyspira hyodysenteriae and Brachyspira pilosicoli in pig faeces. Methods and Results: A multiplex-polymerase chain reaction (M-PCR) was designed to amplify a 655-base pair (bp) portion of the L. intracellularis 16S rRNA gene, a 354-bp portion of the B. hyodysenteriae NADH oxidase gene, and a 823-bp portion of the B. pilosicoli 16S rRNA gene. Specificity was assessed using 80 strains of Brachyspira spp. and 30 other enteric bacteria. Bacterial DNA was extracted from faeces using the QIAamp® DNA Stool Mini Kit. The M-PCR was tested in parallel with culture and/or PCR on 192 faecal samples from eight piggeries. Faeces also were seeded with known cell concentrations of the three pathogenic species, and the limits of detection of the M-PCR tested. The M-PCR was specific, with limits of detection of 102-103 cells of the respective species per gram of faeces. Conclusions: The M-PCR is a rapid, sensitive and specific test for detecting three important enteric bacterial pathogens of pigs. Significance and Impact of the Study: The availability of a new diagnostic M-PCR will allow rapid detection and control of three key porcine enteric pathogens.

Details

Title: Development of a multiplex-PCR for rapid detection of the enteric pathogens Lawsonia intracellularis, Brachyspira hyodysenteriae, and Brachyspira pilosicoli in porcine faeces
Authors/Creators: T. La (Author/Creator) - Murdoch University
A.M. Collins (Author/Creator) - New South Wales Department of Primary Industries
N.D. Phillips (Author/Creator) - Murdoch University
A. Oksa (Author/Creator) - Murdoch University
D.J. Hampson (Author/Creator) - Murdoch University
Publication Details: Letters in Applied Microbiology, Vol.42(3), pp.284-288
Publisher: Blackwell Publishing
Identifiers: 991005542191507891
Copyright: © 2006 The Society for Applied Microbiology
Murdoch Affiliation: School of Veterinary and Biomedical Sciences
Language: English
Resource Type: Journal article

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Collaboration types: Domestic collaboration
Citation topics: 1 Clinical & Life Sciences; 1.248 Sexually Transmitted Infections; 1.248.2445 Lawsonia Intracellularis
Web Of Science research areas: Biotechnology & Applied Microbiology; Microbiology
ESI research areas: Biology & Biochemistry