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Applied and Environmental Microbiology, November 2007, p. 7331-7337, Vol. 73, No. 22
0099-2240/07/$08.00+0     doi:10.1128/AEM.00976-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Multiplex Assay Based on Single-Nucleotide Polymorphisms for Rapid Identification of Brucella Isolates at the Species Level{triangledown} ,{dagger}

Julie C. Scott,{ddagger} Mark S. Koylass, Michael R. Stubberfield,§ and Adrian M. Whatmore*

Department of Statutory and Exotic Bacterial Diseases, Veterinary Laboratories Agency, Addlestone, Surrey KT15 3NB, United Kingdom

Received 1 May 2007/ Accepted 10 September 2007

The genus Brucella includes a number of species that are major animal pathogens worldwide and significant causes of zoonotic infections of humans. Traditional methods of identifying Brucella to the species level can be time-consuming, can be subjective, and can pose a hazard to laboratory personnel in the absence of suitable biocontainment facilities. Using a robust phylogenetic framework, a number of single-nucleotide polymorphisms (SNPs) that define particular species within the genus were identified. These SNPs were used to develop a multiplex SNP detection assay, based on primer extension technology, that can rapidly and unambiguously identify an isolate as a member of one of the six classical Brucella species or as a member of the recently identified marine mammal group.

* Corresponding author. Mailing address: Department of Statutory and Exotic Bacterial Diseases, Veterinary Laboratories Agency, Addlestone, Surrey KT15 3NB, United Kingdom. Phone: 44 1932 357311. Fax: 44 1932 357423. E-mail: a.whatmore{at}vla.defra.gsi.gov.uk

{triangledown} Published ahead of print on 21 September 2007.

{dagger} This paper is dedicated to the memory of Julie C. Scott.

{ddagger} Deceased.

§ Present address: Royal Veterinary College, Hawkshead Lane, Hatfield AL9 7TA, United Kingdom.

Applied and Environmental Microbiology, November 2007, p. 7331-7337, Vol. 73, No. 22
0099-2240/07/$08.00+0     doi:10.1128/AEM.00976-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.





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Copyright © 2007 by the American Society for Microbiology. All rights reserved.