POLYMERASE CHAIN-REACTION FOR IDENTIFICATION OF HUMAN AND PORCINE SPIROCHETES RECOVERED FROM CASES OF INTESTINAL SPIROCHETOSIS

A polymerase chain reaction (PCR) amplification of 16S rDNA was develo ped to identify spirochaetes recovered from cases of intestinal spiroc haetosis in humans and pigs; these bacteria belong to a distinct genet ic group of spirochaetes, with the proposed name 'Anguillina coli'. Th e PCR incorporated a universal eubacterial 16S rDNA sequencing primer (1492r), and a 21-base forward primer designed to include a nucleotide sequence specific for 'A. coli'. The PCR was used to correctly identi fy DNA extracted from 43 isolates of 'A. coli' from humans and pigs, w hilst no product was produced from Escherichia coli, or from other int estinal spirochaetes, including 38 isolates of Serpulina spp., and one each of Treponema succinifaciens and Brachyspira aalborgi. The amplif ication provided a rapid and simple means of identifying DNA from isol ates of 'A. coli', and could be used on boiled whole 'A. coli' cells, with a detection limit equivalent to 2.5 X 10(2) cells. The reaction w as used to detect and identify these spirochaetes from selective agar plates inoculated with stool specimens from infected pigs.