Determining Streptococcus suis serotype from short-read whole-genome sequencing data

BMC Microbiol. 2016 Jul 22;16(1):162. doi: 10.1186/s12866-016-0782-8.

Abstract

Background: Streptococcus suis is divided into 29 serotypes based on a serological reaction against the capsular polysaccharide (CPS). Multiplex PCR tests targeting the cps locus are also used to determine S. suis serotypes, but they cannot differentiate between serotypes 1 and 14, and between serotypes 2 and 1/2. Here, we developed a pipeline permitting in silico serotype determination from whole-genome sequencing (WGS) short-read data that can readily identify all 29 S. suis serotypes.

Results: We sequenced the genomes of 121 strains representing all 29 known S. suis serotypes. We next combined available software into an automated pipeline permitting in silico serotyping of strains by differential alignment of short-read sequencing data to a custom S. suis cps loci database. Strains of serotype pairs 1 and 14, and 2 and 1/2 could be differentiated by a missense mutation in the cpsK gene. We report a 99 % match between coagglutination- and pipeline-determined serotypes for strains in our collection. We used 375 additional S. suis genomes downloaded from the NCBI's Sequence Read Archive (SRA) to validate the pipeline. Validation with SRA WGS data resulted in a 92 % match. Included pipeline subroutines permitted us to assess strain virulence marker content and obtain multilocus sequence typing directly from WGS data.

Conclusions: Our pipeline permits rapid and accurate determination of S. suis serotype, and other lineage information, directly from WGS data. By discriminating between serotypes 1 and 14, and between serotypes 2 and 1/2, our approach solves a three-decade longstanding S. suis typing issue.

Keywords: Serotyping; Short-reads; Streptococcus suis; Whole-genome sequencing.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Capsules
  • Bacterial Proteins
  • Base Sequence
  • DNA, Bacterial / genetics
  • Gene Targeting
  • Genes, Bacterial
  • Genetic Loci
  • Genome, Bacterial
  • Multiplex Polymerase Chain Reaction
  • Polysaccharides, Bacterial / classification
  • Polysaccharides, Bacterial / genetics
  • Polysaccharides, Bacterial / immunology
  • Polysaccharides, Bacterial / isolation & purification
  • Sequence Alignment
  • Sequence Analysis, DNA
  • Serogroup*
  • Serotyping*
  • Streptococcus suis / classification
  • Streptococcus suis / genetics*
  • Streptococcus suis / immunology
  • Streptococcus suis / isolation & purification*
  • Virulence / genetics
  • Virulence Factors

Substances

  • Bacterial Proteins
  • DNA, Bacterial
  • Polysaccharides, Bacterial
  • Virulence Factors