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United States Department of Agriculture

Agricultural Research Service

Title: Development of Species Specific Pcr Tests for Serpulina Hyodysenteriae and S. Pilosicoli Based on Nadh Oxidase Gene Sequence Analysis

Authors
item Jensen, Neil
item Atyeo, Roslyn - MURDOCH UNIV., AUSTRALIA
item Hampson, David - MURDOCH UNIV., AUSTRALIA
item STANTON, THADDEUS

Submitted to: Research Workers in Animal Diseases Conference Proceedings
Publication Type: Abstract Only
Publication Acceptance Date: November 11, 1996
Publication Date: N/A

Technical Abstract: Diarrheal diseases caused by Serpulina sp. affect a broad range of vertebrate hosts. Pathogenic Serpulina sp. include S. hyodysenteriae (swine dysentery in pigs) and S. pilosicoli (intestinal spirochetosis in pigs, dogs, poultry and humans). We have developed PCR primers specific for Serpulina hyodysenteriae (S. hyo) and Serpulina pilosicoli (S. pilo) based on comparative analysis of the NADH oxidase (nox) gene sequences of 16 Serpulina strains representing 5 current or proposed Serpulina species, as well as 2 unclassified Serpulina strains. Previous work has determined that, among the Serpulina species studied, nox gene sequences were variable (less than 94% identity) but conserved within a species. Sequences were aligned using Clustal W 1.60, and regions unique to S. hyo or S. pilo were identified. Oligo 5.0 was used to develop oligonucleotide primers specific for the unique regions of either S. hyo or S. pilo. Serpulina sp. genomic DNA (100 ng) or washed whole cells (10**7-10**8 cells) were used in PCR reactions to assess the specificity of the tests. Amplification products were electrophoresed on 1.2% agarose gels and stained with ethidium bromide. PCR amplification of S. hyo and S. pilo (DNA or cells) resulted in products of 823 and 651 bp in size, respectively. Primers were tested for specificity by running PCR amplifications using other Serpulina sp. DNA or cells. Only S. hyo or S. pilo, depending on which primer pair was used, was amplified. Further study of the sensitivity and specificity of these PCR primers will be carried out with additional Serpulina isolates, and under in vivo conditions in pigs with swine dysentery or intestinal spirochetosis.

Last Modified: 9/29/2014
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