Please use this identifier to cite or link to this item: http://hdl.handle.net/2440/66929
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Type: Journal article
Title: Application of nox-restriction fragment length polymorphism for the differentiation of Brachyspira intestinal spirochetes isolated from pigs and poultry in Australia
Author: Townsend, K.
Giang, V.
Stephens, C.
Scott, P.
Trott, D.
Citation: Journal of Veterinary Diagnostic Investigation, 2005; 17(2):103-109
Publisher: Amer Assoc Veterinary Laboratory Diagnosticians Inc
Issue Date: 2005
ISSN: 1040-6387
1943-4936
Statement of
Responsibility: 
Kirsty M. Townsend, Vo Ngan Giang, Carol Stephens, Paul T. Scott, Darren J. Trott
Abstract: Sixty-nine intestinal spirochetes isolated from pigs and poultry in eastern Australia were selected to evaluate the effectiveness of a species-specific PCR-based restriction fragment length polymorphism (RFLP) analysis of the Brachyspira nox gene. For comparative purposes, all isolates were subjected to species-specific PCRs for the pathogenic species Brachyspira hyodysenteriae and Brachyspira pilosicoli, and selected isolates were examined further by sequence analysis of the nox and 16S ribosomal RNA genes. Modifications to the original nox-RFLP method included direct inoculation of bacterial cells into the amplification mixture and purification of the PCR product, which further optimized the nox-RFLP for use in a veterinary diagnostic laboratory, producing sufficient product for both species identification and future comparisons. Although some novel profiles that prevented definitive identification were observed, the nox-RFLP method successfully classified 45 of 51 (88%) porcine and 15 of 18 (83%) avian isolates into 5 of the 6 recognized species of Brachyspira. This protocol represents a significant improvement over conventional methods currently used in veterinary diagnostic laboratories for rapid specific identification of Brachyspira spp. isolated from both pigs and poultry.
Keywords: Brachyspira; nicotinamide adenine dinucleotide reduced oxidase; PCR; 16S ribosomal DNA sequencing.
Rights: Copyright status unknown
RMID: 0020106846
DOI: 10.1177/104063870501700202
Appears in Collections:Animal and Veterinary Sciences publications

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