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Clinical and Diagnostic Laboratory Immunology, September 1999, p. 760-764, Vol. 6, No. 5
1071-412X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Identification of an IS711 Element Interrupting the wboA Gene of Brucella abortus Vaccine Strain RB51 and a PCR Assay To Distinguish Strain RB51 from Other Brucella Species and Strains

Ramesh Vemulapalli,1 John R. McQuiston,1 Gerhardt G. Schurig,1 Nammalwar Sriranganathan,1 Shirley M. Halling,2 and Stephen M. Boyle1,*

Department of Biomedical Sciences and Pathobiology, Center for Molecular Medicine and Infectious Diseases, Virginia-Maryland Regional College of Veterinary Medicine, Virginia Polytechnic Institute and State University, Blacksburg, Virginia 24061-0342,1 and National Animal Disease Center, ARS, USDA, Ames, Iowa 500112

Received 7 December 1998/Returned for modification 5 April 1999/Accepted 22 June 1999

Brucella abortus vaccine strain RB51 is a natural stable attenuated rough mutant derived from the virulent strain 2308. The genetic mutations that are responsible for the roughness and the attenuation of strain RB51 have not been identified until now. Also, except for an assay based on pulsed-field gel electrophoresis, no other simple method to differentiate strain RB51 from its parent strain 2308 is available. In the present study, we demonstrate that the wboA gene encoding a glycosyltransferase, an enzyme essential for the synthesis of O antigen, is disrupted by an IS711 element in B. abortus vaccine strain RB51. Exploiting this feature, we developed a PCR assay that distinguishes strain RB51 from all other Brucella species and strains tested.

* Corresponding author. Mailing address: Department of Biomedical Sciences and Pathobiology, Center for Molecular Medicine and Infectious Diseases, Virginia-Maryland Regional College of Veterinary Medicine, Virginia Polytechnic Institute and State University, Blacksburg, VA 24061-0342. Phone: (540) 231-4641. Fax: (540) 231-3426. E-mail: smboyle{at}vt.edu.

Clinical and Diagnostic Laboratory Immunology, September 1999, p. 760-764, Vol. 6, No. 5
1071-412X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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