Department of Biology, Faculty of Science, Srinakharinwirot University, Sukhumvit 23, Bangkok 10110, Thailand.
World J Microbiol Biotechnol. 2013 Apr;29(4):721-31. doi: 10.1007/s11274-012-1228-6. Epub 2012 Dec 12.
Monoclonal antibodies (MAbs) specific to Vibrio parahaemolyticus were successfully generated. According to the specificity of V. parahaemolyticus, MAbs can be classified into 5 groups. The MAbs VP-2D and VP-11H were specific to the O2 and O4 groups of V. parahaemolyticus, respectively. The MAb VP-11B reacted with 11 out of 30 isolates of V. parahaemolyticus used in this study. The MAb VP-516 bound to 27 out of 30 isolates of V. parahaemolyticus and cross reacted with all 10 isolates of V. alginolyticus. The MAb VP-618 demonstrated positive reactivity to 29 out of 30 isolates of V. parahaemolyticus and demonstrated slight cross reactivity to 3 out of 30 isolates of V. harveyi. The sensitivity of the MAbs ranged from 10(8) to 10(7) c.f.u. ml(-1) for V. parahaemolyticus obtained from pure cultures and depended on the group of MAbs. However, the detection capability could be improved to be equivalent to that of the PCR technique following pre-incubation of the samples in alkaline peptone water (APW). Using these MAbs along with MAbs specific to V. alginolyticus (VA-165), V. cholerae (VC-63), V. harveyi (VH-9B and VH-20C) and Vibrio spp. (VC-201) from previous studies, V. parahaemolyticus could be identified and differentiated from Vibrio spp. in various seafood samples including shrimp, green mussels, blood clams and oysters by a simple dot blot immunoassay without the requirement for bacterial isolation or biochemical characterization.
成功制备了针对副溶血性弧菌的单克隆抗体（MAbs）。根据副溶血性弧菌的特异性，MAbs 可分为 5 组。MAb VP-2D 和 VP-11H 分别针对副溶血性弧菌的 O2 和 O4 群。MAb VP-11B 与本研究中使用的 30 株副溶血性弧菌分离株中的 11 株反应。MAb VP-516 与 30 株副溶血性弧菌分离株中的 27 株结合，与所有 10 株 Algino 弧菌分离株交叉反应。MAb VP-618 对 30 株副溶血性弧菌分离株中的 29 株表现出阳性反应，对 30 株 Harvey 弧菌分离株中的 3 株表现出轻微交叉反应。MAbs 的灵敏度范围为从纯培养物中获得的副溶血性弧菌 10(8)到 10(7)cfu.ml(-1)，具体取决于 MAb 的组别。然而，通过在碱性蛋白胨水中（APW）对样品进行预孵育，可以将检测能力提高到与 PCR 技术相当的水平。使用这些 MAb 以及之前针对 Algino 弧菌（VA-165）、霍乱弧菌（VC-63）、Harvey 弧菌（VH-9B 和 VH-20C）和 Vibrio spp.（VC-201）的 MAb，可以通过简单的点印迹免疫测定法在各种海鲜样品（包括虾、贻贝、血蛤和牡蛎）中识别和区分副溶血性弧菌，而无需进行细菌分离或生化鉴定。
