The annealing site of each primer was identified by BLASTing the primer’s sequence against publically accessible CP673451 solubility dmso S. pneumoniae genomic sequences available through the National Center for Biotechnology Information [28, 29]. These results identified where each primer annealed

relative to the typing region, and whether the sequencing resulting from the primer was able to consistently cover the required region. This full process was replicated twice for each primer set and each test isolate to confirm the AZD5582 reproducibility of the observations. Acknowledgements The authors would like to acknowledge the Canadian Immunization Monitoring Program Active Investigators for collecting the S. pneumoniae isolates that made this project possible. The Canadian Immunization Monitoring Program Active is a national surveillance initiative managed by the Canadian Pediatric Society (CPS) and conducted by the IMPACT investigators on behalf of the Public Health Agency of Canada’s (PHAC) Centre for Immunization and Respiratory Infectious Diseases. The authors would also like to acknowledge Cynthia Bishop for providing

her guidance during this investigation and her permission to reference the personal communications between herself and the author’s research team. Funding Funding for collection of the pneumococcal isolates used in this ON-01910 clinical trial study was provided by an unrestricted grant to CPS from Wyeth Pharmaceuticals (1991–2005), and the PHAC (2005–2009). Funding to support the laboratory analysis was provided by Pfizer Canada through an investigator-initiated research grant in aid to Dr. James D. Kellner. Tolmetin Electronic supplementary material Additional file 1: Table S1: S. pneumoniae strains sequence typed with alternative MLST primers. (DOC 105 KB) References 1. Maiden MC, Bygraves JA, Feil E, Morelli G, Russell JE, Urwin R, Zhang Q, Zhou J, Zurth K, Caugant DA, Feavers IM, Achtman M, Spratt BG: Multilocus sequence

typing: a portable approach to the identification of clones within populations of pathogenic microorganisms. Proc Natl Acad Sci U S A 1998,95(6):3140–3145.PubMedCentralPubMedCrossRef 2. Urwin R, Maiden MCJ: Multi-locus sequence typing: a tool for global epidemiology. Trends Microbiol 2003,11(10):479–487.PubMedCrossRef 3. Bentley SD, Aanensen DM, Mavroidi A, Saunders D, Rabbinowitsch E, Collins M, Donohoe K, Harris D, Murphy L, Quail MA, Samuel G, Skovsted IC, Kaltoft MS, Barrell B, Reeves PR, Parkhill J, Spratt BG: Genetic analysis of the capsular biosynthetic locus from All 90 pneumococcal serotypes. PLoS Genet 2006,2(3):e31.PubMedCentralPubMedCrossRef 4.