日本鳗鲡体表溃疡病病原菌的分离、鉴定及单克隆抗体制备  

Isolation and identification of bacteriosis pathogen from Anguilla japonica ulceration of skin and preparation of its monoclonal antibody

作　　者：樊海平[1] 吴斌[1] 曾占壮[1] 钟全福[1] 张新艳[1] 辛志明[1] 林煜[1] 严乾临 

机构地区：[1]福建省淡水水产研究所,福建福州350002 [2]福建省动物卫生监督所,福建福州350003

出　　处：《中国水产科学》2009年第2期295-302,共8页Journal of Fishery Sciences of China

基　　金：福建省海洋与渔业科技项目(闽海渔科2007008号)

摘　　要：从日本鳗鲡(Anguilla japonica)体表溃疡病的肝脏分离到一株优势细菌JS70322NA,经人工感染证实为引起日本鳗鲡体表溃疡病的病原菌。通过形态学观察、API20E细菌鉴定试剂盒鉴定、生理生化试验和16SrRNA特异性基因分析,鉴定该菌为嗜水气单胞菌(Aeromonas hydrophila)。JS70322NA为典型的β-溶血,血清型为O∶9。以甲醛灭活的JS70322NA为抗原免疫Balb/c小鼠,利用淋巴细胞杂交瘤技术,制备出3株抗嗜水气单胞菌的单克隆抗体,分别命名为:1A5、1B8、1F3;其腹水效价分别达1.0×10-6、1.0×10-5、1.0×10-6;细胞亚型分别为IgG2a、IgM、IgG2a;3株单克隆抗体与温和气单胞菌(Aeromonas sobria)、豚鼠气单胞菌(Aeromonas caviae)、鳗弧菌(Vibrio anguillarum)等6种水产常见致病菌无交叉反应,特异强,可应用于日本鳗鲡嗜水气单胞菌特异性检测。Bacterium strain JS70322NA,isolated from diseased Anguilla japonica in March 2007,was confirmed as the causative pathogen associated with ulceration of skin by artificial infection test.Strain JS70322NA was identified as Aeromonas hydrophila by physiological,API 20E biochemical items and 16S rRNA tests.The strain wasβ-hemolysis, and was serotyped as O∶9 with the same agglutination of Ah9617.Strain JS70322NA was inactivated by formaldehyde to prepare antigens for the immunization of Balb/c mice.Three strains of hybridomas,secreting McAb against A.hydrophila,were obtained by means of hybridoma technique.The titers of three McAbs in ascites were 1.0×10^-6, 1.0×10 -5 and 1.0×10 -6 ,respectively;the isotype of the McAbs were IgG2a,IgM and IgG2a,respectively,and three strains of McAbs had no cross reactions with 6 aquatic pathogenic bacteria.The results indicated that the McAbs specially against A.hydrophila could be used for rapid detection and isolation of infected Japanese eel.

关 键 词：日本鳗鲡 嗜水气单胞菌 细菌鉴定 溶血活性 血清型 单克隆抗体 

分 类 号：S96[农业科学—水产养殖]