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. 2003 Feb;41(2):601-7.
doi: 10.1128/JCM.41.2.601-607.2003.

Evaluation of semiautomated multiplex PCR assay for determination of Streptococcus pneumoniae serotypes and serogroups

Elliot R Lawrence  1 , David B Griffiths, Siobhán A Martin, Robert C George, Lucinda M C Hall
Affiliations

Evaluation of semiautomated multiplex PCR assay for determination of Streptococcus pneumoniae serotypes and serogroups

Elliot R Lawrence et al. J Clin Microbiol. 2003 Feb.

Abstract

A semiautomated method for the determination of five serotypes and three serogroups in Streptococcus pneumoniae was developed. Primers specific for serotypes 1, 3, 14, 19F, and 23F and serogroups 6, 19, and 23 were combined in three multiplex PCRs. Products were separated by capillary electrophoresis with a 7-min run time, and a serotype or serogroup was assigned on the basis of fragment size. The method was used to test 93 clinical isolates, and all isolates of the serotypes concerned were correctly detected. The strategy would allow the detection of multiple serotypes in a single sample. Detection of additional serotypes could be included as capsule locus sequences become available.

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Figures

FIG. 1.
FIG. 1.
Agarose gel (2%) electrophoresis showing the multiplex PCR assay results for four clinical isolates (isolates PN1010, PN1008, PN1001, and PN1106). Lane M, 50-bp DNA ladder (Gibco-BRL); lanes 1 to 4 are multiplex PCRs: 3M (serotypes 3, 14, and 23F), 19M (serotypes 1 and 19F), GPM (serogroups 6, 19, and 23), and CPSA (pneumococcal capsule gene cpsA), respectively.
See this image and copyright information in PMC

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