应用间接荧光抗体技术检测林蛙嗜水气单胞菌  

Detection of a Bacterium Aeromonas hydrophila in Wood Frog by Indirect Fluorescent Antibody Technique

作　　者：白雪梅[1] 李太元[1] 杨兴[1] 李艳茹[1] 孙丹丹[1] 董丹[1] 

机构地区：[1]延边大学农学院,吉林龙井133400

出　　处：《水产科学》2010年第10期613-615,共3页Fisheries Science

基　　金：吉林省科技发展计划项目(20050703-4)

摘　　要：以感染嗜水气单胞菌的林蛙各组织脏器为抗原,以热灭活的嗜水气单胞菌为免疫原,免疫家兔制备兔抗嗜水气单胞菌血清为第一抗体,用异硫氰酸荧光素(FITC)标记的羊抗兔IgG作为二抗,建立了间接荧光抗体试验方法。分别采用2.5%戊二醛5 min、甲醇10 min、冷丙酮1 h 3种固定方法固定抗原,然后滴加不同稀释倍数的兔抗嗜水气单胞菌血清、羊抗兔IgG-FITC,于荧光显微镜下观察。结果表明,2.5%戊二醛固定腹水5 min,兔抗血清抗体效价为1∶80,荧光二抗的最佳稀释倍数为1∶4000。A new indirect fluorescent antibody test（IFAT） method was established to identify a bacterium Aeromonas hydrophila in wood frog quickly and accurately.In the IFAT the organs and tissues taken from wood frog infected with the bacterium were used as antigen,heat-inactivated Aeromonas hydrophila as the immunogen,the bacterium in the serum from anti-immune rabbit as the first antibody,and caprine anti-rabbit IgG labeled with fluorescein isothiocyanate（FITC） as the second antibody.In the identification procedure,the antigen was firstly taken and fixed by 2.5% glutaraldehyde for 5 min,methanol for 10 min,and cold acetone for 1 h in turn.Then the rabbit anti-Aeromonas hydrophila serum with different dilution and caprine anti-rabbit IgG-FITC was added drop by drop into the fixed antigen.Finally,the processed antigen was observed under fluorescence microscope.Results showed that the first antibody titer was 1：80 and the optimal dilution for the secondary antibody labeled with FITC was 1∶4000 under antigen of ascites fixed by the first kind（2.5% glutaraldehyde for 5 min）.

关 键 词：荧光抗体技术 林蛙 嗜水气单胞菌 

分 类 号：S966.35[农业科学—水产养殖]