Direct and Rapid Detection by PCR of <i>Erysipelothrix</i> sp. DNAs Prepared from Bacterial Strains and Animal Tissues

<jats:title>ABSTRACT</jats:title> <jats:p> A PCR method for rapid screening of <jats:italic>Erysipelothrix</jats:italic> spp. in the slaughterhouse was carried out by using four species-specific sets of oligonucleotide primers after initial amplification with the primer set MO101-MO102, which amplifies the 16S rRNA sequences of all four <jats:italic>Erysipelothrix</jats:italic> species. The DNA sequences coding for the rRNA gene cluster, including 16S rRNA, 23S rRNA, and the noncoding region downstream of 5S rRNA, were determined in order to design primers for the species-specific PCR detection system. The homology among the 4.5-kb DNA sequences of the rRNA genes of <jats:italic>Erysipelothrix rhusiopathiae</jats:italic> serovar 2 (DNA Data Bank of Japan accession no. <jats:ext-link xmlns:xlink="http://www.w3.org/1999/xlink" ext-link-type="gen" xlink:href="AB019247" xlink:type="simple">AB019247</jats:ext-link> ), <jats:italic>E. tonsillarum</jats:italic> serovar 7 (accession no. <jats:ext-link xmlns:xlink="http://www.w3.org/1999/xlink" ext-link-type="gen" xlink:href="AB019248" xlink:type="simple">AB019248</jats:ext-link> ), <jats:italic>E. rhusiopathiae</jats:italic> serovar 13 (accession no. <jats:ext-link xmlns:xlink="http://www.w3.org/1999/xlink" ext-link-type="gen" xlink:href="AB019249" xlink:type="simple">AB019249</jats:ext-link> ), and <jats:italic>E. rhusiopathiae</jats:italic> serovar 18 (accession no. <jats:ext-link xmlns:xlink="http://www.w3.org/1999/xlink" ext-link-type="gen" xlink:href="AB019250" xlink:type="simple">AB019250</jats:ext-link> ) ranged from 96.0 to 98.4%. The PCR amplifications were specific and were able to distinguish the DNAs from each of the four <jats:italic>Erysipelothrix</jats:italic> species. The results of PCR tests performed directly with tissue specimens from diseased animals were compared with the results of cultivation tests, and the PCR tests were completed within 5 h. The test with this species-specific system based on PCR amplification with the DNA sequences coding for the rRNA gene cluster was an accurate, easy-to-read screening method for rapid diagnosis of <jats:italic>Erysipelothrix</jats:italic> sp. infection in the slaughterhouse. </jats:p>