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Journal of Clinical Microbiology, July 2006, p. 2398-2403, Vol. 44, No. 7
0095-1137/06/$08.00+0     doi:10.1128/JCM.02236-05
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Molecular Characterization of Nontypeable Group B Streptococcus

Srinivas V. Ramaswamy,1 Patricia Ferrieri,2 Aurea E. Flores,2 and Lawrence C. Paoletti1*

Channing Laboratory, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02115,1 Departments of Laboratory Medicine and Pathology and Pediatrics, University of Minnesota Medical School, Minneapolis, Minnesota 554552

Received 25 October 2005/ Returned for modification 14 December 2005/ Accepted 29 March 2006

Traditionally, the capsular polysaccharide (CPS) antigen has been used to distinguish between the nine known serotypes of group B streptococcus (GBS) by classical antibody-antigen reactions. In this study, we used PCR for all CPSs and selected protein antigens, multilocus sequencing typing (MLST), and pulsed-field gel electrophoresis (PFGE) to molecularly characterize 92 clinical isolates identified as nontypeable (NT) by CPS-specific antibody-antigen reactivity. The PCR and MLST were performed on blinded, randomly numbered isolates. All isolates contained the cfb gene coding for CAMP factor. While most (56.5%) contained a single CPS-specific gene, 40 isolates contained either two or three CPS-specific genes. Type V CPS-specific gene was present in 66% of the isolates, and all serotypes except types IV, VII, and VIII were represented. Most (44.5%) of the isolates contained a single protein antigen gene (bca, bac, rib, alp1, or alp3), and the remaining isolates had multiple protein antigen genes. Of the 61 isolates that had the V CPS-specific gene, 48 (78.6%) had the alp3 gene. PFGE analysis classified the isolates into 21 profile groups, while MLST analysis divided the isolates into 16 sequence types. Forty-two (69%) of 61 isolates with the V CPS-specific gene were in PFGE profile group 4; 41 of these 42 were sequence type 1 by MLST. These data shed new light on the antigenic complexity of NT GBS isolates, information that can be valuable in the formulation of an effective GBS vaccine.

* Corresponding author. Mailing address: Channing Laboratory, Brigham and Women's Hospital, Harvard Medical School, 221 Longwood Avenue, Boston, MA 02115. Phone: (617) 525-7878. Fax: (617) 525-2682. E-mail: lpaoletti{at}channing.harvard.edu.

Journal of Clinical Microbiology, July 2006, p. 2398-2403, Vol. 44, No. 7
0095-1137/06/$08.00+0     doi:10.1128/JCM.02236-05
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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