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Volume 54, Issue 4

A molecular-capsular-type prediction system for 90 serotypes using partial sequencing and - or -specific PCR Free

Abstract

In a previous study, a molecular capsular type (MCT) prediction system for 51 serotypes was developed based on a combination of partial – sequencing and serotype(s)/group(s)-specific PCR. In this study, another 169 isolates were added to the existing database of 427 isolates, including representatives of all 39 serotypes not previously studied. In addition to the authors’ own limited sequence data for all 90 serotypes, – sequence data published by the capsular loci-sequencing group (http://www.sanger.ac.uk/Projects/S_pneumoniae/CPS/) at the Sanger Institute or available from GenBank were incorporated into the database. All serotypes, except 25A, were represented by at least two isolates. The number of sequence types identified was 138, of which 110 corresponded to single conventional serotypes (CSs); of these, 57 were represented by two or more isolates. Twenty-six sequence types were shared by between two and four CSs. To resolve these shared – sequence types and increase the discriminatory power of our system, the genes encoding the capsular polysaccharide flippase () and polymerase () were annotated and 24 new serotype(s)/group(s)-specific PCRs targeting and two targeting were designed. Using both – sequencing and / PCR, MCT correctly predicted the CSs of 516 (73 %) and the serogroup of an additional 155 (22 %) of the 708 isolates evaluated. For 5 % of isolates, MCT could not distinguish between members of five serotype pairs (37 isolates) containing members of different serogroups. Although further study of the relationship between MCT and CS is needed, this system now allows serotype or serogroup identification of 95 % of isolates.

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2005-04-01
2024-04-19
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