Sequetyping: Serotyping Streptococcus pneumoniae by a Single PCR Sequencing Strategy
| Autor: | Bruno Pichon, Stephen H. Gillespie, Marcus H. Y. Leung, Kathrin Freystatter, Kevin Bryson, Bambos M. Charalambous, Giles Edwards |
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| Rok vydání: | 2012 |
| Předmět: |
DNA
Bacterial Microbiology (medical) Serotype Molecular Sequence Data Biology medicine.disease_cause Polymerase Chain Reaction Sensitivity and Specificity Pneumococcal Infections Pneumococcal conjugate vaccine law.invention law Conjugate vaccine Multiplex polymerase chain reaction Streptococcus pneumoniae medicine Humans Serotyping Conserved Sequence Polymerase chain reaction DNA Primers Computational Biology Reproducibility of Results Bacteriology Sequence Analysis DNA medicine.disease Vaccine efficacy Virology Molecular Typing Pneumococcal infections medicine.drug |
| Zdroj: | Journal of Clinical Microbiology. 50:2419-2427 |
| ISSN: | 1098-660X 0095-1137 |
| Popis: | The introduction of pneumococcal conjugate vaccines necessitates continued monitoring of circulating strains to assess vaccine efficacy and replacement serotypes. Conventional serological methods are costly, labor-intensive, and prone to misidentification, while current DNA-based methods have limited serotype coverage requiring multiple PCR primers. In this study, a computer algorithm was developed to interrogate the capsulation locus ( cps ) of vaccine serotypes to locate primer pairs in conserved regions that border variable regions and could differentiate between serotypes. In silico analysis of cps from 92 serotypes indicated that a primer pair spanning the regulatory gene cpsB could putatively amplify 84 serotypes and differentiate 46. This primer set was specific to Streptococcus pneumoniae , with no amplification observed for other species, including S. mitis , S. oralis , and S. pseudopneumoniae . One hundred thirty-eight pneumococcal strains covering 48 serotypes were tested. Of 23 vaccine serotypes included in the study, most (19/22, 86%) were identified correctly at least to the serogroup level, including all of the 13-valent conjugate vaccine and other replacement serotypes. Reproducibility was demonstrated by the correct sequetyping of different strains of a serotype. This novel sequence-based method employing a single PCR primer pair is cost-effective and simple. Furthermore, it has the potential to identify new serotypes that may evolve in the future. |
| Databáze: | OpenAIRE |
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