引用本文:	池信才,王　军,鄢庆枇,苏永全.大黄鱼病原副溶血弧菌单克隆抗体制备及其应用[J].海洋科学,2007,31(8):1-5.

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大黄鱼病原副溶血弧菌单克隆抗体制备及其应用
池信才¹, 王　军¹, 鄢庆枇², 苏永全¹
1.厦门大学海洋环境学院;2.集美大学水产学院

摘要:

用甲醛灭活副溶血弧菌(Vibrio parahaemolyticus)和溶藻弧菌(V.alginolyticus)制成免疫原免疫BALB/c小鼠,利用淋巴细胞杂交瘤技术,获得1株特异的针对副溶血弧菌的单克隆抗体,命名为D6F3H5。腹水及培养上清液抗体的ELISA效价分别为:1∶5 120和1∶1 280,该单克隆抗体与其它细菌没有明显的交叉反应。利用该单克隆抗体和兔抗副溶血弧菌多克隆抗体,建立了检测副溶血弧菌的双抗体夹心ELISA方法。该方法对副溶血弧菌的最小检出量为5×10⁴个/mL。用双抗体夹心ELISA检测大黄鱼(Pseudosciaena crocea)样品,14尾患病大黄鱼中有11尾检出副溶血弧菌,而10尾健康大黄鱼都没有检出副溶血弧菌。由此可见,本试验制备的高特异性的抗副溶血弧菌单克隆抗体,可以用于患病大黄鱼副溶血弧菌的快速诊断。

关键词: 大黄鱼(Pseudosciaena crocea) 副溶血弧菌(Vibrio parahaemolyticus) 单克隆抗体

DOI：

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基金项目:国家863计划项目(2004AA623010,2002AA603021);厦门市科技局项目

Preparation and applications of monoclonal antibody against the pathogenic Vibrio parahaemolyticus from Pseudosciaena crocea

Abstract:

Vibrio parahaemolyticus and V.alginolyticus were inactivated by formaldehyde to prepare antigens for the immunizing of BALB/c mice. 1 strain of hybridoma(D6F3H5), secreting McAb against V.parahaemolyticus ,was obtained by means of hybridoma technique. The ELISA titers of McAb in ascites and culture liquid were 1∶5 120 and 1∶1 280, respectively. The McAb had no cross reaction to 12 strains of other bacteria. Sandwich ELISA was established to detect V.parahaemolyticus based on the McAb and rabbit against V.parahaemolyticus multi-clone antibody. The lowest detectable concentration of V.parahaemolyticus detected by sandwich ELISA was 5×10⁴ cells/mL. Sandwich ELISA was used for detecting V.parahaemolyticus in large yellow croacker, Pseudosciaena crocea, V.parahaemolyticus was detected from 11 fish among 14 diseased P.crocea, no V.parahaemolyticus was detected from 10 healthy P.crocea. The results indicated that the McAb specially against V.parahaemolyticus could be used for the rapid diagnosis of diseased P.crocea.

Key words: Pseudosciaena crocea Vibrio parahaemolyticus monoclonal antibody