The Medical Microbiology Program, Graduate School, Chulalongkorn University, Bangkok, Thailand.
Department of Veterinary Public Health, Faculty of Veterinary Sciences, Chulalongkorn University, Bangkok, Thailand.
PLoS One. 2024 Apr 3;19(4):e0299691. doi: 10.1371/journal.pone.0299691. eCollection 2024.
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.
猪链球菌血清 2 型(SS2)是一种重要的猪病原体,可引起猪和人类的疾病。为了快速鉴定 SS2,开发了一种基于重链可变区抗体(VH)的新型乳胶凝集试验(LAT)。首先,在大肠杆菌中表达了来自噬菌体展示文库的可溶性 47B3 VH 抗体片段,该文库在 C 末端设计了半胱氨酸残基。然后,将纯化的蛋白质温和还原形成单体可溶性 47B3 VH,随后通过位点特异性缀合将其涂覆在乳胶珠上。所得 VH 涂覆的珠粒与 SS2 产生良好的凝集反应。LAT 能够区分 SS2 血清型 1/2,因为它们具有一些共同的糖残基,并且与其他 SS2 血清型或其他相关细菌没有交叉反应。检测灵敏度高达 1.85x106 个细胞。LAT 在 4°C 下至少稳定六个月,而不会失去活性。据我们所知,这是第一种基于 VH 抗体片段的 LAT,可以替代传统的基于抗体的 LAT,其中 VH 是最受欢迎的重组抗体。