Group B streptococcus (GBS), Streptococcus agalactiae MLST

• Definition of MLST for Streptococcus agalactiae
• GBS MLST loci
• GBS MLST database
• References cited in GBS MLST database
• mlstdb software

Image curtesy of Seth Pincus, LSU Health Sciences Center and Fred Hayes, NIAID Rocky Mountain Laboratories

Definition of MLST for Streptococcus agalactiae

MLST has been developed for Streptococcus agalactiae (group B streptococci, GBS) and an initial database of the allelic profiles of 152 isolates obtained from cases of serious neonatal and adult invasive disease and asymptomatic genitourinary carriage* has been set up. Many of the GBS capsular serotypes are represented in this set.

* Jones N, Bohnsack JF, Takahashi S, Oliver K, Chan MS, Kunst F, Glaser P, Rusniok C, Crook DC, Harding R, Bisharat N, Spratt BG. A Multilocus Sequence Typing System for Group B Streptococcus. Journal of Clinical Microbiology 2003; 41(6):2530-6.

The GBS MLST scheme has been developed by Nicola Jones (Nuffield Department of Clinical Laboratory Sciences, John Radcliffe Hospital, Oxford, OX3 9DU, United Kingdom) in the laboratory of Brian Spratt (now at the Department of Infectious Disease Epidemiology, Faculty of Medicine, Imperial College, St. Mary’s Hospital, London W2 1PG, United Kingdom, [email protected]) together with Dr Derrick Crook (Nuffield Department of Clinical Laboratory Sciences, John Radcliffe Hospital, Oxford, OX3 9DU, United Kingdom, [email protected]).

For further details about the MLST scheme please contact Nicola Jones. Nicola Jones ([email protected]) For queries about the web site or database contact Man-Suen Chan ([email protected])

GBS MLST loci

Internal fragments (400-500 bp) of the alcohol dehydrogenase (adhP), phenylalanyl tRNA synthetase (PheS), glutamine transporter protein (atr), glutamine synthetase (glnA), serine dehydratase (sdhA), glucose kinase (glcK) and transketolase (tkt) genes were sequenced.

Oligonucleotide primers for GBS MLST

Obtaining an allelic profile and comparing your strains with those in our database

Access database

The allelic profile of a GBS strain is obtained by sequencing the internal fragments of the seven house-keeping genes. The sequence of each fragment must be obtained on both strands, and they must be 100% accurate, since even a single error may convert a known allele into a novel allele. The sequences have to be trimmed so that they correspond exactly to the region that we use to define the alleles. The sequences of the seven loci from a typical GBS can be obtained and can be used to ensure that your sequences have been trimmed correctly. You then need to access our databases, which involves a simple registration process that allows us to inform you of new developments by e-mail. Select the GBS database, and the multiple locus query, followed by submit. You then cut and paste your seven sequences into the corresponding boxes and submit them. Alternatively you can assign your alleles one locus at a time by selecting the single locus query option and, having obtained the alleles at the seven loci, you can select allelic profile query and enter the seven integers. The software will check that the sequences are the correct length and that they do not contain any unrecognised characters. A check is also made to see if the submitted sequence is at least 70% similar to another allele at that locus (in case you have cut and pasted a sequence into the wrong box). If your sequence is different from any of the alleles in our database you should submit the sequence traces to Nicola Jones ([email protected]) who will check your data, and provide you with a new allele number, and add your new allele to our database. After submitting the seven sequences, you will obtain the allelic profile of your isolate.

References cited in GBS MLST database

1. Bohnsack, J. F., S. Takahashi, S. R. Detrick, L. R. Pelinka, L. L. Hammitt, A. A. Aly, A. A. Whiting, and E. E. Adderson. 2001. Phylogenetic classification of serotype III group B streptococci on the basis of hylB gene analysis and DNA sequences specific to restriction digest pattern type III-3. J Infect Dis 183:1694-7.

2. Bohnsack, J. F., S. Takahashi, L. Hammitt, D. V. Miller, A. A. Aly, and E. E. Adderson. 2000. Genetic polymorphisms of group B streptococcus scpB alter functional activity of a cell-associated peptidase that inactivates C5a. Infect Immun 68:5018-25.

3. Bohnsack, J. F., A. A. Whiting, R. D. Bradford, B. K. Van Frank, S. Takahashi, and E. E. Adderson. 2002. Long-range mapping of the Streptococcus agalactiae phylogenetic lineage restriction digest pattern type III-3 reveals clustering of virulence genes. Infect Immun 70:134-9.

4. Chaffin, D. O., S. B. Beres, H. H. Yim, and C. E. Rubens. 2000. The serotype of type Ia and III group B streptococci is determined by the polymerase gene within the polycistronic capsule operon. J Bacteriol 182:4466-77.

5. Coffey, T. J., M. C. Enright, M. Daniels, J. K. Morona, R. Morona, W. Hryniewicz, J. C. Paton, and B. G. Spratt. 1998. Recombinational exchanges at the capsular polysaccharide biosynthetic locus lead to frequent serotype changes among natural isolates of Streptococcus pneumoniae. Mol Microbiol 27:73-83.

6. Dingle, K. E., F. M. Colles, D. R. Wareing, R. Ure, A. J. Fox, F. E. Bolton, H. J. Bootsma, R. J. Willems, R. Urwin, and M. C. Maiden. 2001. Multilocus sequence typing system for Campylobacter jejuni. J Clin Microbiol 39:14-23.

7. Embley, T. M. 1991. The linear PCR reaction: a simple and robust method for sequencing amplified rRNA genes. Lett Appl Microbiol 13:171-4.

8. Enright, M. C., B. G. Spratt, A. Kalia, J. H. Cross, and D. E. Bessen. 2001. Multilocus sequence typing of Streptococcus pyogenes and the relationships between emm type and clone. Infect Immun 69:2416-27.

9. Glaser, P., C. Rusniok, C. Buchrieser, F. Chevalier, L. Frangeul, T. Msadek, M. Zouine, E. Couve, L. Lalioui, C. Poyart, P. Trieu Cuot, and F. Kunst. 2002. Genome sequence of Streptococcus agalactiae, a pathogen causing invasive neonatal disease. Molecular Microbiology 45:1.

10. Hauge, M., C. Jespersgaard, K. Poulsen, and M. Kilian. 1996. Population structure of Streptococcus agalactiae reveals an association between specific evolutionary lineages and putative virulence factors but not disease. Infect Immun 64:919-25.

11. Jolley, K. A., E. J. Feil, M. S. Chan, and M. C. Maiden. 2001. Sequence type analysis and recombinational tests (START). Bioinformatics 17:1230-1.

12. Maiden, M. C., J. A. Bygraves, E. Feil, G. Morelli, J. E. Russell, R. Urwin, Q. Zhang, J. Zhou, K. Zurth, D. A. Caugant, I. M. Feavers, M. Achtman, and B. G. Spratt. 1998. Multilocus sequence typing: a portable approach to the identification of clones within populations of pathogenic microorganisms. Proc Natl Acad Sci U S A 95:3140-5.

13. Musser, J. M., S. J. Mattingly, R. Quentin, A. Goudeau, and R. K. Selander. 1989. Identification of a high-virulence clone of type III Streptococcus agalactiae (group B Streptococcus) causing invasive neonatal disease. Proc Natl Acad Sci U S A 86:4731-5.

14. PHLS. 2002. Incidence of group B streptococcal disease in infants aged less than 90 days. CDR weekly 12 (16):3.

15. Quentin, R., H. Huet, F. S. Wang, P. Geslin, A. Goudeau, and R. K. Selander. 1995. Characterization of Streptococcus agalactiae strains by multilocus enzyme genotype and serotype: identification of multiple virulent clone families that cause invasive neonatal disease. J Clin Microbiol 33:2576-81.

16. Schuchat, A. 1999. Group B streptococcus. Lancet 353:51-6.

17. Staden, R. 1996. The Staden sequence analysis package. Mol Biotechnol 5:233-41.

18. Tamura, G. S., M. Herndon, J. Przekwas, C. E. Rubens, P. Ferrieri, and S. L. Hillier. 2000. Analysis of restriction fragment length polymorphisms of the insertion sequence IS1381 in group B Streptococci. J Infect Dis 181:364-8.

19. Tettelin, H., V. Masignani, M. J. Cieslewicz, J. A. Eisen, S. Peterson, M. R. Wessels, I. T. Paulsen, K. E. Nelson, I. Margarit, T. D. Read, L. C. Madoff, A. M. Wolf, M. J. Beanan, L. M. Brinkac, S. C. Daugherty, R. T. DeBoy, A. S. Durkin, J. F. Kolonay, R. Madupu, M. R. Lewis, D. Radune, N. B. Fedorova, D. Scanlan, H. Khouri, S. Mulligan, H. A. Carty, R. T. Cline, S. E. Van Aken, J. Gill, M. Scarselli, M. Mora, E. T. Iacobini, C. Brettoni, G. Galli, M. Mariani, F. Vegni, D. Maione, D. Rinaudo, R. Rappuoli, J. L. Telford, D. L. Kasper, G. Grandi, and C. M. Fraser. 2002. Complete genome sequence and comparative genomic analysis of an emerging human pathogen, serotype V Streptococcus agalactiae. Proc Natl Acad Sci U S A.

20. Wilder-Smith, E., K. M. Chow, R. Kay, M. Ip, and N. Tee. 2000. Group B streptococcal meningitis in adults: recent increase in Southeast Asia. Aust N Z J Med 30:462-5. [back to top]