抗布鲁氏菌单克隆抗体的研制及cELISA方法的建立

Development of monoclonal antibodies to Brucellaand validation of competitive enzyme-linked immunosorbent assay

利用杂交瘤技术筛选出3株抗布鲁氏菌的特异性单克隆抗体5C10、4D2和3E4;利用单抗5C10建立了检测血清中布鲁氏菌抗体的竞争ELISA(cELISA)方法。用该方法对320份牛血清样品和147份绵羊血清样品进行了检测,并与商品化布鲁氏菌病试剂盒进行了比较。结果显示,用建立的cELISA方法和商品化布鲁氏菌病试剂盒对320份牛血清样品和147份绵羊血清样品的检测符合率分别为95.6%和92.5%。用补体结合试验(CFT)进一步检测差异结果的样品(25份),证明建立的cELISA与CFT的吻合率更高(21/25),而商品化布鲁氏菌病试剂盒与CFT的吻合率较低(4/25)。结果表明,本研究建立的用于检测布鲁氏菌抗体的cELISA方法更特异敏感,在布鲁氏菌病根除计划中将具有重要的应用价值。

In order to accurate detection of brucella from animals,three strains of monoclonal antibodies(MAbs)against Brucella,which was named as 5C10,4D2,and 3E4,were obtained by using hybridoma technique.With one of the MAbs 5C10,a competitive enzyme-linked immunosorbent assay(cELISA)for the serological detection of brucellosis was developed.Three hundred and twenty bovine and 147 ovine sera were used for the comparison of the detection effect of cELISA and the commercial kit.From the result,the agreement rates of the cELISA with the commercial kit were 95.6%and 92.5%for the bovine and ovine sera,respectively.The discrepant samples(25)were confirmed by the complement fixation test(CFT).We found that coincidence rate of the developed cELISA with CFT was higher(21/25)than the commercial kit(4/25).These results suggested that our cELISA for serological detection is more specific and sensitive than the commercial kit.It will be very useful in eradication program for brucellosis.