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Journal of Clinical Microbiology, December 1999, p. 4093-4098, Vol. 37, No. 12
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

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

Kouichi Takeshi,1,* Souichi Makino,2 Tetsuya Ikeda,1 Noriko Takada,3 Atsushi Nakashiro,4 Kazunori Nakanishi,4 Keiji Oguma,5 Yoshinobu Katoh,1 Hiroyuki Sunagawa,1 and Tohru Ohyama1

Department of Food Science, Hokkaido Institute of Public Health, Sapporo 060,1 Department of Veterinary Microbiology, Obihiro University of Agriculture and Veterinary Medicine, Obihiro 080,2 Kurume Health Center of Fukuoka Prefectural Government, Kurume 830,3 Meat Inspection Office of Hokkaido Prefectural Government, Sapporo 060,4 and Department of Bacteriology, Okayama University Medical School, Okayama 700,5 Japan

Received 19 March 1999/Returned for modification 13 July 1999/Accepted 24 August 1999

A PCR method for rapid screening of Erysipelothrix 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 Erysipelothrix 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 Erysipelothrix rhusiopathiae serovar 2 (DNA Data Bank of Japan accession no. AB019247), E. tonsillarum serovar 7 (accession no. AB019248), E. rhusiopathiae serovar 13 (accession no. AB019249), and E. rhusiopathiae serovar 18 (accession no. AB019250) ranged from 96.0 to 98.4%. The PCR amplifications were specific and were able to distinguish the DNAs from each of the four Erysipelothrix 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 Erysipelothrix sp. infection in the slaughterhouse.

* Corresponding author. Mailing address: Department of Food Science, Hokkaido Institute of Public Health, Kita 19, Nishi 12, Kita-ku, Sapporo, Hokkaido 060-0819, Japan. Phone: 001-81-11-747-2211. Fax: 001-81-11-736-9476. E-mail: takeshi{at}iph.pref.hokkaido.jp.

Journal of Clinical Microbiology, December 1999, p. 4093-4098, Vol. 37, No. 12
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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