title: Use of pyrosequencing to differentiate Streptococcus pneumoniae serotypes 6A and 6B.
abstract: Accurate serotyping of Streptococcus pneumoniae remains important to monitor the changes in seroepidemiology of the organism over time. Though several PCR-based systems have been developed for this purpose, the cross-reactivity within serogroups often limits discrimination between types. All serogroup 6 isolates can be identified using a multiplex PCR system; however, due to the high sequence homology between the cps-6B and cps-6A loci, serotypes 6A and 6B cannot be differentiated by this method. We describe the use of pyrosequencing to reliably differentiate between serotypes 6A and 6B using a previously described single nucleotide polymorphism at codon 195 of the cps locus wciP gene. We observed complete concordance between capsular serotyping results and wciP pyrosequencing among 210 isolates examined, indicating that pyrosequencing is a rapid and accurate technique for deducing serotypes 6A and 6B.
authors.affiliation: Bacterial and Mycotic Diseases, Respiratory Diseases Branch, Atlanta, GA 30333, USA.
authors.forename: Rekha
authors.initials: R
authors.lastname: Pai
authors.forename: Josef
authors.initials: J
authors.lastname: Limor
authors.forename: Bernard
authors.initials: B
authors.lastname: Beall
mesh_terms: D016667:Bacterial Capsules / Q000235:genetics / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics*; D015373:Bacterial Typing Techniques; D001483:Base Sequence; D006602:Hexosyltransferases / Q000737:chemistry / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*; D017422:Sequence Analysis, DNA; D012703:Serotyping; D013296:Streptococcus pneumoniae / Q000145:classification* / Q000201:enzymology / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D006602:Hexosyltransferases; C088697:rhamnosyltransferase 1