Streptococcus suis serotyping by a new multiplex PCR

Kerdsin, Anusak; Akeda, Yukihiro; Hatrongjit, Rujirat; Detchawna, Unchaya; Sekizaki, Tsutomu; Hamada, Shigeyuki; Gottschalk, Marcelo; Oishi, Kazunori

doi:10.1099/jmm.0.069757-0

Cited by 85 publications

(100 citation statements)

References 22 publications

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“…has been developed in recent years (13,14,34); the specific cps genes are planned for use in typing. This gene cluster strongly supports the results of agglutination assays and avoids the false positivity of naked-eye observation in the agglutination test.…”

Section: Discussionmentioning

confidence: 99%

“…Phylogenetic analyses of the cps gene cluster, conserved Wzy polymerase, Wzx flippase, and glycosyltransferase are all taken as important means of classifying a novel serotype (12). Multiplex PCR assays against the specific genes of the cps clusters have also been developed to identify serotypes in S. suis (13)(14)(15).From March to May 2013, strain CZ130302 caused an outbreak of streptococcosis in piglets at multiple large-scale pig farms in Jiangsu Province, China. This pathogenic bacterium induced meningitis in 30-day-old piglets, with a total morbidity rate of 25% to 35%.…”

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confidence: 99%

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confidence: 99%

“…The serotyping primers were designed based on the sequences of capsule loci (cps), which were described in earlier reports (13,14). Using the multiplex PCR system, four reactions were developed to detect all serotypes of S. suis.…”

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Novel Variant Serotype of Streptococcus suis Isolated from Piglets with Meningitis

Pan

Wang³

et al. 2015

Appl Environ Microbiol

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cStreptococcus suis is an emerging zoonotic pathogen causing severe infections in pigs and humans. In previous studies, 33 serotypes of S. suis have been identified using serum agglutination. Here, we describe a novel S. suis strain, CZ130302, isolated from an outbreak of acute piglet meningitis in eastern China. Strong pathogenicity of meningitis caused by strain CZ130302 was reproduced in the BALB/c mouse model. The strain showed a high fatality rate (8/10), higher than those for known virulent serotype 2 strains P1/7 (1/10) and 9801 (2/10). Cell adhesion assay results with bEnd.3 and HEp2 cells showed that CZ130302 was significantly close to P1/7 and 9801. Both the agglutination test and its complementary test showed that strain CZ130302 had no strong cross-reaction with the other 33 S. suis serotypes. The multiplex PCR assays revealed no specified bands for all four sets used to detect the other 33 serotypes. In addition, genetic analysis of the whole cps gene clusters of all serotypes was performed in this study. The results of comparative genomics showed that the cps gene cluster of CZ130302, which was not previously reported, showed no homology to the gene sequences of the other strains. Especially, the wzy, wzx, and acetyltransferase genes of strain CZ130302 are phylogenetically distinct from strains of the other 33 serotypes. Therefore, this study suggested that strain CZ130302 represents a novel variant serotype of S. suis (designated serotype Chz) which has a high potential to be virulent and associated with meningitis in animals. Streptococcus suis causes meningitis and septicemia in pigs and is also known as a zoonotic agent (1). Human infections of S. suis were first reported in Denmark in 1968 (2). Since then, this pathogen has spread all over the world. The human Streptococcus suis was epidemic in most Europe countries (3, 4), as well as in Asian countries, such as Vietnam and Thailand (5-7). In China, two outbreaks of human streptococcosis have occurred, affecting more than 100 people and causing 39 deaths (8). More and more S. suis infections from China, Thailand, Hong Kong, Taiwan, and Singapore have been reported, which indicates that S. suis has been an important cause of adult meningitis, endocarditis, septicemia, and arthritis in Asia (9).The serotyping of S. suis isolates rests on the basis of the antigenicity of their capsular polysaccharides (CPs); 35 serotypes have been identified by agglutination tests (10). With the development of sequence analysis of 16S rRNA and cpn60 genes in S. suis, the original S. suis serotypes 32 and 34 were reclassified as Streptococcus orisratti (11). Phylogenetic analyses of the cps gene cluster, conserved Wzy polymerase, Wzx flippase, and glycosyltransferase are all taken as important means of classifying a novel serotype (12). Multiplex PCR assays against the specific genes of the cps clusters have also been developed to identify serotypes in S. suis (13)(14)(15).From March to May 2013, strain CZ130302 caused an outbreak of streptococcosis in piglets at ...

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Section: Discussionmentioning

confidence: 99%

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confidence: 99%

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Novel Variant Serotype of Streptococcus suis Isolated from Piglets with Meningitis

Pan

Wang³

et al. 2015

Appl Environ Microbiol

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“…The problems associated with the production of the antisera and testing have led to the development of different molecular approaches, of which multiplex PCR (mPCR) with amplification of serotype-specific cps genes are an attractive alternative to the existing serological tests. In fact, three mPCR assays were developed to identify S. suis serotypes (Kerdsin et al, 2014;Liu et al, 2013;Okura et al, 2014). However, two or three sets of mPCR amplification are needed when using these assays.…”

Section: Introductionmentioning

confidence: 99%

Simultaneous detection of 33 Streptococcus suis serotypes using the luminex xTAG® assay™

Bai

Liu

et al. 2015

Journal of Microbiological Methods

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The establishment and application of a one‐step multiplex real‐time polymerase chain reaction assay for the detection of Streptococcus suis, Streptococcus suis serotype 2, and Glaesserella parasuis

Xin,

Wang,

et al. 2023

Animal Research and One Health

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This article aims to establish a multiplex real‐time polymerase chain reaction (PCR) assay for the simultaneous detection of Streptococcus suis (SS), Streptococcus suis serotype 2 (SS2), and Glaesserella parasuis (GPS). In this study, three pairs of primers and three probes were designed based on the specific sequences of SS (gdh), SS2 (cps2j), and GPS (infB). The results showed that the assay was not cross‐reacted with other swine pathogens (Escherichia coli, Pasteurella multocida, Staphylococcus aureus, Streptococcus agalactiae, Streptococcus pneumoniae, Actinobacillus pleuropneumoniae, Mycoplasma hyopneumoniae, and Enterococcus faecalis; Streptococcus pyogenes). 108 to 102 copies/μL showed the R2 values for SS, SS2, and GPS were 0.999, 0.992, and 0.990, respectively. The multiplex real‐time PCR efficiency was 93.816% for gdh, 105.260% for cps2j, and 93.175% for infB. The sensitivity result showed that SS, SS2, and GPS could be detected at 10 copies/μL. The repeatability result showed that intra‐assay and inter‐assay coefficients of variation of SS, SS2, and GPS were <2%. The best cutoff values for SS, SS2, and GPS were determined from ROC curves to be 35.085, 35.620, and 34.940, respectively. Areas under the curve were 0.943, 0.968, and 0.958. In total, 88 clinical samples were analyzed. The results indicated positive rates of 11.364% (10/88) for SS, 20.455% (18/88) for SS2, and 18.182% (16/88) for GPS. In conclusion, the developed one‐step multiplex real‐time PCR assay may be a valuable tool for the early detection of the SS, SS2 and, GPS with high specificity and sensitivity.

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Streptococcus suis serotyping by a new multiplex PCR

Abstract: A multiplex PCR was developed to detect all true serotypes of Streptococcus suis. This multiplex PCR was composed of four reaction sets. The first set identified nine serotypes (serotypes 1/2,

Cited by 85 publications

References 22 publications

Novel Variant Serotype of Streptococcus suis Isolated from Piglets with Meningitis

Novel Variant Serotype of Streptococcus suis Isolated from Piglets with Meningitis

Simultaneous detection of 33 Streptococcus suis serotypes using the luminex xTAG® assay™

The establishment and application of a one‐step multiplex real‐time polymerase chain reaction assay for the detection of Streptococcus suis, Streptococcus suis serotype 2, and Glaesserella parasuis

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