Development of Multiplex PCR Assays for the Identification of the 33 Serotypes of Streptococcus suis

University of Helsinki, Finland
PLoS ONE (Impact Factor: 3.23). 08/2013; 8(8):e72070. DOI: 10.1371/journal.pone.0072070
Source: PubMed


Streptococcussuis is an important zoonotic agent causing severe diseases in pigs and humans. To date, 33 serotypes of S. suis have been identified based on antigenic differences in the capsular polysaccharide. The capsular polysaccharide synthesis (cps) locus encodes proteins/enzymes that are responsible for capsular production and variation in the capsule structures are the basis of S. suis serotyping. Multiplex and/or simplex PCR assays have been developed for 15 serotypes based on serotype-specific genes in the cps gene cluster. In this study, we developed a set of multiplex PCR (mPCR) assays to identify the 33 currently known S. suis serotypes. To identify serotype-specific genes for mPCR, the entire genomes of reference strains for the 33 serotypes were sequenced using whole genome high-throughput sequencing, and the cps gene clusters from these strains were identified and compared. We developed a set of 4 mPCR assays based on the polysaccharide polymerase gene wzy, one of the serotype-specific genes. The assays can identify all serotypes except for two pairs of serotypes: 1 and 14, and 2 and 1/2, which have no serotype-specific genes between them. The first assay identifies 12 serotypes (serotypes 1 to 10, 1/2, and 14) that are the most frequently isolated from diseased pigs and patients; the second identifies 10 serotypes (serotypes 11 to 21 except 14); the third identifies the remaining 11 serotypes (serotypes 22 to 31, and 33); and the fourth identifies a new cps cluster of S. suis discovered in this study in 16 isolates that agglutinated with antisera for serotypes 29 and 21. The multiplex PCR assays developed in this study provide a rapid and specific method for molecular serotyping of S. suis.

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    • "Our multiplex PCR assay was developed to identify the 29 currently known Streptococcus suis serotypes. A multiplex PCR identifying 33 serotypes of Streptococcus suis was recently developed; however, the former Streptococcus suis serotypes 20, 22, 26 and 33 were still included in that assay (Liu et al., 2013). Our multiplex PCR assay targets true Streptococcus suis serotypes only. "
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    ABSTRACT: A multiplex polymerase chain reaction (PCR) was developed to detect all true serotypes of Streptococcus suis. This multiplex PCR was composed of 4 reaction sets. The first set identified 9 serotypes (serotypes 1/2, 1, 2, 3, 7, 9, 11, 14 and 16), the second set identified 8 serotypes (serotypes 4, 5, 8, 12, 18, 19, 24 and 25), the third set identified 7 serotypes (serotypes 6, 10, 13, 15, 17, 23 and 31), and the last set identified 5 serotypes (serotypes 21, 27, 28, 29 and 30). This assay correctly detected serotypes 2, 5, 14 and 24 in human isolates, and serotypes 1, 2, 1/2, 3, 4, 5, 6, 7, 8, 9, 11, 12, 14, 15, 16, 17, 19, 24, 28 and 31 in pig isolates from Thailand. No cross-reaction was observed with other bacterial species. Multiplex PCR was able to simultaneously allow amplifying DNA mixture of reference S.suis serotypes. This assay should be useful for serotype surveillance of human and pig isolates of S. suis.
    Journal of Medical Microbiology 04/2014; 63(Pt_6). DOI:10.1099/jmm.0.069757-0 · 2.25 Impact Factor
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    ABSTRACT: We developed a practical and easy two-step multiplex PCR assay to aid in serotyping of Streptococcus suis. The assay accurately typed almost all of the serotype reference strains and field isolates of various serotypes and also identified the genotypes of capsular polysaccharide synthesis gene clusters of some serologically nontypeable strains.
    Journal of clinical microbiology 02/2014; 52(5). DOI:10.1128/JCM.03411-13 · 3.99 Impact Factor
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    ABSTRACT: Streptococcus suis (S. suis) is a family of pathogenic gram-positive bacterial strains that represents a primary health problem in the swine industry worldwide. S. suis is also an emerging zoonotic pathogen that causes severe human infections clinically featuring with varied diseases/syndromes (such as meningitis, septicemia, and arthritis). Over the past few decades, continued efforts have made significant progress toward better understanding this zoonotic infectious entity, contributing in part to the elucidation of the molecular mechanism underlying its high pathogenicity. This review is aimed at presenting an updated overview of this pathogen from the perspective of molecular epidemiology, clinical diagnosis and typing, virulence mechanism, and protective antigens contributing to its zoonosis.
    Virulence 03/2014; 5(4). DOI:10.4161/viru.28595 · 4.22 Impact Factor
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