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Clinical and Diagnostic Laboratory Immunology, September 2000, p. 835-839, Vol. 7, No. 5
1071-412X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

An IS711 Element Downstream of the bp26 Gene Is a Specific Marker of Brucella spp. Isolated from Marine Mammals

Axel Cloeckaert,* Maggy Grayon, and Olivier Grepinet

Laboratoire de Pathologie Infectieuse et Immunologie, Institut National de la Recherche Agronomique, 37380 Nouzilly, France

Received 23 March 2000/Returned for modification 22 May 2000/Accepted 6 June 2000

DNA polymorphism of the bp26 gene, coding for a diagnostic protein antigen for brucellosis, was assessed by PCR and restriction fragment length polymorphism analysis using primers to amplify the bp26 gene with its flanking regions. Surprisingly, whereas PCR performed on DNA of the reference strains of the six recognized Brucella species produced a product of the expected size (1,029 bp), PCR performed on DNA of three representative strains from marine mammals (from a seal, a dolphin, and a porpoise) produced a larger product, of about 1,900 bp. Nucleotide sequencing of the 1,900-bp PCR products revealed the presence of an insertion sequence, IS711, downstream of the bp26 gene and adjacent to a Bru-RS1 element previously described as being a hot spot for IS711 insertion. PCR performed on a large number of field strains from different geographic origins and from marine mammal isolates indicated that the occurrence of an IS711 element downstream of the bp26 gene was a feature specific to the marine mammal Brucella strains. Thus, this PCR assay is able to differentiate Brucella terrestrial isolates from marine mammal isolates and could be applied for diagnostic purposes.

* Corresponding author. Present address: Institut National de la Recherche Agronomique, Station de Pathologie Aviaire et Parasitologie, 37380 Nouzilly, France. Phone: (33) 2 47 42 77 50. Fax: (33) 2 47 42 77 74. E-mail: cloeckae{at}tours.inra.fr.

Clinical and Diagnostic Laboratory Immunology, September 2000, p. 835-839, Vol. 7, No. 5
1071-412X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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