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Journal of Clinical Microbiology, October 1999, p. 3417-3420, Vol. 37, No. 10
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Restriction Fragment Length Polymorphisms of Virulence Plasmids in Rhodococcus equi

Shinji Takai,1,* Masato Shoda,1 Yukako Sasaki,1 Shiro Tsubaki,1 Guillaume Fortier,2 Stephane Pronost,2 Karim Rahal,3 Teotimo Becu,4 Angela Begg,5 Glenn Browning,6 Vivian M. Nicholson,7 and John F. Prescott7

Kitasato University, Towada, Aomori 034-8628, Japan1; Laboratoire Frank Duncombe, Caen, Calvados,2 and École Nationale Vétérinaire de Lyon,3 France; Clínica Equina S.R.L., Capitán Sarmiento, Provincia de Buenos Aires, Argentina4; Scone Veterinary Diagnostic Laboratory, Scone,5 and Veterinary Preclinical Center, The University of Melbourne, Parkville, Victoria 3052,6 Australia; and Ontario Veterinary College, University of Guelph, Ontario, Canada7

Received 23 December 1998/Returned for modification 27 April 1999/Accepted 19 July 1999

Virulent Rhodococcus equi, which is a well-known cause of pyogranulomatous pneumonia in foals, possesses a large plasmid encoding virulence-associated 15- to 17-kDa antigens. Foal and soil isolates from five countries---Argentina, Australia, Canada, France, and Japan---were investigated for the presence of 15- to 17-kDa antigens by colony blotting, using the monoclonal antibody 10G5, and the gene coding for 15- to 17-kDa antigens by PCR. Plasmid DNAs extracted from positive isolates were digested with restriction endonucleases BamHI, EcoRI, EcoT22I, and HindIII, and the digestion patterns that resulted divided the plasmids of virulent isolates into five closely related types. Three of the five types had already been reported in Canadian and Japanese isolates, and the two new types had been found in French and Japanese isolates. Therefore, we tentatively designated these five types 85-kb type I (pREAT701), 85-kb type II (a new type), 87-kb type I (EcoRI and BamHI type 2 [V. M. Nicholson and J. F. Prescott, J. Clin. Microbiol. 35:738-740, 1997]), 87-kb type II (a new type), and 90-kb (pREL1) plasmids. The 85-kb type I plasmid was found in isolates from Argentina, Australia, Canada, and France. Plasmid 87-kb type I was isolated in specimens from Argentina, Canada, and France. The 85-kb type II plasmid appeared in isolates from France. On the other hand, plasmids 87-kb type II and 90-kb were found only in isolates from Japan. These results revealed geographic differences in the distribution of the virulence plasmids found in the five countries and suggested that the restriction fragment length polymorphism of virulence plasmids might be useful to elucidate the molecular epidemiology of virulent R. equi in the world.

* Corresponding author. Mailing address: Department of Animal Hygiene, School of Veterinary Medicine and Animal Sciences, Kitasato University, Towada, Aomori 034-8628, Japan. Phone: 81-167-23-4371. Fax: 81-176-23-8703. E-mail: takai{at}vmas.kitasato-u.ac.jp.

Journal of Clinical Microbiology, October 1999, p. 3417-3420, Vol. 37, No. 10
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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