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Journal of Clinical Microbiology, September 2005, p. 4820-4822, Vol. 43, No. 9
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.9.4820-4822.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Use of Pyrosequencing To Differentiate Streptococcus pneumoniae Serotypes 6A and 6B

Rekha Pai,1 Josef Limor,2 and Bernard Beall1*

Bacterial and Mycotic Diseases, Respiratory Diseases Branch,1 Scientific Resources Program, Biotechnology Core Facility Branch, Centers for Disease Control and Prevention, Atlanta, Georgia2

Received 29 April 2005/ Returned for modification 30 May 2005/ Accepted 8 June 2005

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.

* Corresponding author. Mailing address: CDC Respiratory Diseases Branch, Mailstop C02, 1600 Clifton Rd., NE, Atlanta, GA 30333. Phone: (404) 639-1237. Fax: (404) 639-4215. E-mail: BBEALL{at}CDC.GOV.

Journal of Clinical Microbiology, September 2005, p. 4820-4822, Vol. 43, No. 9
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.9.4820-4822.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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