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Novel PCR-Restriction Fragment Length Polymorphism Method for Determining Serotypes or Serogroups of Streptococcus pneumoniae Isolates

Centre for Medical Microbiology, University College London, Royal Free Campus, Rowland Hill St., London NW3 2PF,¹ Specialist and Reference Microbiology Division, Health Protection Agency, 67, Colindale Avenue, London NW9 5HT, United Kingdom²

Received 4 August 2004/ Returned for modification 27 September 2004/ Accepted 7 February 2005

Serotyping Streptococcus pneumoniae is a technique generally confined to reference laboratories, as purchasing pneumococcal antisera is a huge investment. Many attempts have been made to modify serological agglutination techniques to make them more accessible, and more recently developments in serotyping have focused on molecular techniques. This paper describes a PCR assay which amplifies the entire capsulation locus between dexB and aliA. Amplicons are digested to produce serotype-specific patterns. We have shown, using 81 epidemiologically unrelated strains representing 46 different serotypes, that the patterns correlate with a 90 to 100% similarity range for the same serotype or serogroup. Prospective testing of 73 isolates of unknown serotype confirmed reliable serotype attribution, and serotype profiles are reproducible on repeated testing. Once our database contains all 90 serotypes, this technique should be fully portable, cost-effective, and useful in any laboratory with sufficient molecular experience.

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