Development of a latex agglutination test based on VH antibody fragment for detection of Streptococcus suis serotype 2

PLoS One. 2024 Apr 3;19(4):e0299691. doi: 10.1371/journal.pone.0299691. eCollection 2024.

Abstract

Streptococcus suis serotype 2 (SS2) is an important porcine pathogen that causes diseases in both swine and human. For rapid SS2 identification, a novel latex agglutination test (LAT) based on heavy-chain variable domain antibody (VH) was developed. Firstly, the soluble 47B3 VH antibody fragment from a phage display library, in which cysteine residues were engineered at the C-terminus, was expressed in Escherichia coli. The purified protein was then gently reduced to form monomeric soluble 47B3 VH subsequently used to coat with latex beads by means of site-specific conjugation. The resulting VH-coated beads gave a good agglutination reaction with SS2. The LAT was able to distinguish S. suis serotype 2 from serotype 1/2, which shares some common sugar residues, and showed no cross-reaction with other serotypes of S. suis or other related bacteria. The detection sensitivity was found to be as high as 1.85x106 cells. The LAT was stable at 4°C for at least six months without loss of activity. To the best of our knowledge, this is the first LAT based on a VH antibody fragment that can be considered as an alternative for conventional antibody-based LAT where VHs are the most favored recombinant antibody.

MeSH terms

  • Animals
  • Escherichia coli / genetics
  • Humans
  • Immunoglobulin Fragments
  • Latex Fixation Tests / methods
  • Recombinant Proteins / genetics
  • Serogroup
  • Streptococcal Infections* / microbiology
  • Streptococcus suis*
  • Swine
  • Swine Diseases* / microbiology

Substances

  • Immunoglobulin Fragments
  • Recombinant Proteins

Grants and funding

All reagents used in this project was supported by Ratchadapisek Research Funds from Chulalongkorn University, grant number CU_GR_63_17_33_06 and fund from Food Risk Hub, Research Unit of Chulalongkorn University, Bangkok, Thailand.