Bead-based DNA assay for the detection od Streptococcus suis in tonsillar specimens of pigs

F.J. van der Wal, H.J. Wisselink, J. Bergervoet, M. de Weerdt, R.P. Achterberg

Research output: Chapter in Book/Report/Conference proceedingAbstractAcademic

Abstract

Streptococcus suis is an important swine pathogen in nearly all countries with an extensive pig industry. In pigs it is associated with meningitis, arthritis, endocarditis, septicemia, pneumonia and sudden death. In certain parts of Asia S. suis is a common cause of bacterial meningitis in humans. Attempts to control S. suis in pig farming are hampered by the lack of sensitive diagnostic tools and effective vaccines. To provide a basis for adequate control measures, a bead-based suspension array was developed that targets six genes using multiplex PCR followed by target specific primer extension (TSPE) and subsequent hybridisation to beads. Assays were designed using AlleleID and Primer- Plex on consensus sequences of four serotype specific capsular polysaccharide (cps) genes (cps1I, cps2J, cps7H, cps9H), the extracellular virulence factor EF (epf) and a general S. suis marker (glutamate dehydrogenase; gdh). The proof-of-principle of the assay was demonstrated with chromosomal DNA of reference strains: each PCR was capable of producing the desired amplicon, also when performed in a sixplex reaction. Upon hybridisation of the TSPE products, all multiplex PCR products produced signals on the appropriate beads. This assay will be evaluated with field samples, using DNA isolated from tonsillar specimens of pigs. Ultimately, this expandable assay may be applicable for studying epidemiology and transmission, and could contribute to efforts aimed at control and eradication of S. suis in pig farming, thereby contributing to human health.
LanguageEnglish
Title of host publicationMed-Vet-Net Association International Scientific Conference DTU, Lyngby, Denmark, 24-25 June 2013: Abstract Book. Lyngby, 2013
Pages27
Publication statusPublished - 2013
EventMed-Vet-Net Association International Scientific Conference DTU - Lyngby, Denmark
Duration: 24 Jun 201325 Jun 2013

Conference

ConferenceMed-Vet-Net Association International Scientific Conference DTU
CountryDenmark
CityLyngby
Period24/06/1325/06/13

Fingerprint

Streptococcus suis
swine
DNA
assays
hybridization
farming systems
endocarditis
glutamate dehydrogenase
consensus sequence
meningitis
septicemia
arthritis
pneumonia
human health
control methods
serotypes
virulence
polysaccharides
genes
death

Cite this

van der Wal, F. J., Wisselink, H. J., Bergervoet, J., de Weerdt, M., & Achterberg, R. P. (2013). Bead-based DNA assay for the detection od Streptococcus suis in tonsillar specimens of pigs. In Med-Vet-Net Association International Scientific Conference DTU, Lyngby, Denmark, 24-25 June 2013: Abstract Book. Lyngby, 2013 (pp. 27)
van der Wal, F.J. ; Wisselink, H.J. ; Bergervoet, J. ; de Weerdt, M. ; Achterberg, R.P. / Bead-based DNA assay for the detection od Streptococcus suis in tonsillar specimens of pigs. Med-Vet-Net Association International Scientific Conference DTU, Lyngby, Denmark, 24-25 June 2013: Abstract Book. Lyngby, 2013. 2013. pp. 27
@inbook{c705947321774e1b98cc2449b8209f29,
title = "Bead-based DNA assay for the detection od Streptococcus suis in tonsillar specimens of pigs",
abstract = "Streptococcus suis is an important swine pathogen in nearly all countries with an extensive pig industry. In pigs it is associated with meningitis, arthritis, endocarditis, septicemia, pneumonia and sudden death. In certain parts of Asia S. suis is a common cause of bacterial meningitis in humans. Attempts to control S. suis in pig farming are hampered by the lack of sensitive diagnostic tools and effective vaccines. To provide a basis for adequate control measures, a bead-based suspension array was developed that targets six genes using multiplex PCR followed by target specific primer extension (TSPE) and subsequent hybridisation to beads. Assays were designed using AlleleID and Primer- Plex on consensus sequences of four serotype specific capsular polysaccharide (cps) genes (cps1I, cps2J, cps7H, cps9H), the extracellular virulence factor EF (epf) and a general S. suis marker (glutamate dehydrogenase; gdh). The proof-of-principle of the assay was demonstrated with chromosomal DNA of reference strains: each PCR was capable of producing the desired amplicon, also when performed in a sixplex reaction. Upon hybridisation of the TSPE products, all multiplex PCR products produced signals on the appropriate beads. This assay will be evaluated with field samples, using DNA isolated from tonsillar specimens of pigs. Ultimately, this expandable assay may be applicable for studying epidemiology and transmission, and could contribute to efforts aimed at control and eradication of S. suis in pig farming, thereby contributing to human health.",
author = "{van der Wal}, F.J. and H.J. Wisselink and J. Bergervoet and {de Weerdt}, M. and R.P. Achterberg",
year = "2013",
language = "English",
pages = "27",
booktitle = "Med-Vet-Net Association International Scientific Conference DTU, Lyngby, Denmark, 24-25 June 2013: Abstract Book. Lyngby, 2013",

}

van der Wal, FJ, Wisselink, HJ, Bergervoet, J, de Weerdt, M & Achterberg, RP 2013, Bead-based DNA assay for the detection od Streptococcus suis in tonsillar specimens of pigs. in Med-Vet-Net Association International Scientific Conference DTU, Lyngby, Denmark, 24-25 June 2013: Abstract Book. Lyngby, 2013. pp. 27, Med-Vet-Net Association International Scientific Conference DTU, Lyngby, Denmark, 24/06/13.

Bead-based DNA assay for the detection od Streptococcus suis in tonsillar specimens of pigs. / van der Wal, F.J.; Wisselink, H.J.; Bergervoet, J.; de Weerdt, M.; Achterberg, R.P.

Med-Vet-Net Association International Scientific Conference DTU, Lyngby, Denmark, 24-25 June 2013: Abstract Book. Lyngby, 2013. 2013. p. 27.

Research output: Chapter in Book/Report/Conference proceedingAbstractAcademic

TY - CHAP

T1 - Bead-based DNA assay for the detection od Streptococcus suis in tonsillar specimens of pigs

AU - van der Wal, F.J.

AU - Wisselink, H.J.

AU - Bergervoet, J.

AU - de Weerdt, M.

AU - Achterberg, R.P.

PY - 2013

Y1 - 2013

N2 - Streptococcus suis is an important swine pathogen in nearly all countries with an extensive pig industry. In pigs it is associated with meningitis, arthritis, endocarditis, septicemia, pneumonia and sudden death. In certain parts of Asia S. suis is a common cause of bacterial meningitis in humans. Attempts to control S. suis in pig farming are hampered by the lack of sensitive diagnostic tools and effective vaccines. To provide a basis for adequate control measures, a bead-based suspension array was developed that targets six genes using multiplex PCR followed by target specific primer extension (TSPE) and subsequent hybridisation to beads. Assays were designed using AlleleID and Primer- Plex on consensus sequences of four serotype specific capsular polysaccharide (cps) genes (cps1I, cps2J, cps7H, cps9H), the extracellular virulence factor EF (epf) and a general S. suis marker (glutamate dehydrogenase; gdh). The proof-of-principle of the assay was demonstrated with chromosomal DNA of reference strains: each PCR was capable of producing the desired amplicon, also when performed in a sixplex reaction. Upon hybridisation of the TSPE products, all multiplex PCR products produced signals on the appropriate beads. This assay will be evaluated with field samples, using DNA isolated from tonsillar specimens of pigs. Ultimately, this expandable assay may be applicable for studying epidemiology and transmission, and could contribute to efforts aimed at control and eradication of S. suis in pig farming, thereby contributing to human health.

AB - Streptococcus suis is an important swine pathogen in nearly all countries with an extensive pig industry. In pigs it is associated with meningitis, arthritis, endocarditis, septicemia, pneumonia and sudden death. In certain parts of Asia S. suis is a common cause of bacterial meningitis in humans. Attempts to control S. suis in pig farming are hampered by the lack of sensitive diagnostic tools and effective vaccines. To provide a basis for adequate control measures, a bead-based suspension array was developed that targets six genes using multiplex PCR followed by target specific primer extension (TSPE) and subsequent hybridisation to beads. Assays were designed using AlleleID and Primer- Plex on consensus sequences of four serotype specific capsular polysaccharide (cps) genes (cps1I, cps2J, cps7H, cps9H), the extracellular virulence factor EF (epf) and a general S. suis marker (glutamate dehydrogenase; gdh). The proof-of-principle of the assay was demonstrated with chromosomal DNA of reference strains: each PCR was capable of producing the desired amplicon, also when performed in a sixplex reaction. Upon hybridisation of the TSPE products, all multiplex PCR products produced signals on the appropriate beads. This assay will be evaluated with field samples, using DNA isolated from tonsillar specimens of pigs. Ultimately, this expandable assay may be applicable for studying epidemiology and transmission, and could contribute to efforts aimed at control and eradication of S. suis in pig farming, thereby contributing to human health.

M3 - Abstract

SP - 27

BT - Med-Vet-Net Association International Scientific Conference DTU, Lyngby, Denmark, 24-25 June 2013: Abstract Book. Lyngby, 2013

ER -

van der Wal FJ, Wisselink HJ, Bergervoet J, de Weerdt M, Achterberg RP. Bead-based DNA assay for the detection od Streptococcus suis in tonsillar specimens of pigs. In Med-Vet-Net Association International Scientific Conference DTU, Lyngby, Denmark, 24-25 June 2013: Abstract Book. Lyngby, 2013. 2013. p. 27