DEVELOPMENT OF NEW DIAGNOSTIC PCR TESTS FOR THE DETECTION OF SERPULINA SPECIES IN CLINICAL SAMPLES (COMBINED BIOLOGICAL SCIENCES MTG., PERTH, AUSTRALIA)

Authors: ATYEO, ROSLYN - MURDOCH UNIV., AUSTRALIA; JENSEN, NEIL - 3625-30-15; Stanton, Thaddeus; HAMPSON, DAVID - MURDOCH UNIV., AUSTRALIA

Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: 9/15/1996
Publication Date: N/A
Citation: N/A

Interpretive Summary:

Technical Abstract: Spirochaetes inhabiting the large intestine are required to tolerate very low amounts of oxygen in their environment, and appear to grow more efficiently in culture when low levels (1%) of oxygen are incorporated into media. The ability of spirochaetes and other colonic bacteria to utilize low amounts of oxygen has been attributed to the presence of NADH oxidase enzymes, which have been identified in all Serpulina species described to date, as well as in a variety of other colonic flora. The NADH oxidase gene in 14 strains of a variety of species from the genus Serpulina, and the type strain of Brachyspira aalborgii, was sequenced. This data was aligned and used to generate PCR tests for the Serpulina species S. hyodysenteriae, "S. intermedia", "S. murdochii", S. innocens, and S. pilosicoli. Of these, S. hyodysenteriae, "S. intermedia" and S. pilosicoli have been associated with intestinal disease. The tests were optimized by screening purified DNA from 61 different strains of spirochaetes. Most tests were specific for the species for which they were designed, except for S. pilosicoli, where DNA was not amplified from all strains. These tests will be important tools in rapid identification of spirochaetes present in clinical samples, and will assist to identify pathogenic species. This is the first PCR reported for "S. intermedia", a proposed new species which is emerging as an important pathogen of poultry.