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Development of a high-throughput MassARRAY-based single assay for the characterization of Streptococcus suis species

Surasak Jiemsup1, Kamonwan Lunha2, Wiyada Chumpol2

  • 1National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), Phahonyothin Road, Khlong Luang, Pathum Thani, 12120, Thailand. surasak@biotec.or.th.

Scientific Reports
|March 6, 2025
PubMed

Insights

A new MassARRAY assay accurately identifies Streptococcus suis and all 29 serotypes. This rapid method aids surveillance and control of this important swine pathogen and zoonotic agent.

Area of Science:

  • Veterinary Microbiology
  • Molecular Diagnostics
  • Public Health

Background:

  • Streptococcus suis is a major cause of disease in pigs and humans globally.
  • Accurate identification and serotyping of S. suis are vital for disease surveillance and control.
  • Current methods can be time-consuming or lack comprehensive coverage of all serotypes.

Purpose of the Study:

  • To develop a novel, single MassARRAY-based assay for simultaneous S. suis species identification and differentiation of all 29 serotypes.
  • To provide a rapid, accurate, and efficient diagnostic tool for S. suis surveillance.

Main Methods:

  • Targeted amplification of glutamate dehydrogenase (gdh) and recombination/repair protein (recN) genes for species identification.
  • Targeted amplification of capsular polysaccharide (cps) genes and analysis of single nucleotide polymorphisms (SNPs) for serotyping.
  • Validation using genomic DNA from 105 whole-genome sequencing (WGS)-confirmed isolates and 143 field isolates.

Main Results:

  • The MassARRAY assay achieved 100% specificity and sensitivity for species identification and serotyping in WGS-validated isolates.
  • Evaluation with field isolates showed perfect agreement with WGS for species identification (100% sensitivity/specificity) and high agreement for serotyping (99.15% sensitivity, 100% specificity, κ=0.98).
  • The assay demonstrated a low limit of detection (1-10 pg gDNA) and accurately distinguished specific serotype pairs based on SNPs.

Conclusions:

  • The developed MassARRAY assay is a highly specific and sensitive tool for simultaneous S. suis species identification and serotyping.
  • This novel assay offers a valuable advancement for routine diagnostics, surveillance, and outbreak investigations of S. suis.
  • The assay's efficiency and accuracy support improved management of S. suis infections in both animal and human health.
Keywords:
Streptococcus suisMALDI-TOFMass spectrometryMassARRAY assaySerotyping

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