双抗体夹心酶联免疫吸附试验检测布氏菌抗原的实验研究  

Experimental Study on double antibodies sandwich Enzymelinked Immurosorbent Assay for detection of Brucella antigens

作　　者：赵鸿雁[1] 鲁齐发[1] 

机构地区：[1]中国疾病预防控制中心传染病预防控制所,北京102206

出　　处：《中国地方病防治》2004年第3期151-153,共3页Chinese Journal of Control of Endemic Diseases

摘　　要：目的　建立检测布氏菌抗原特异、敏感和快速的试验方法。方法　在制备高滴度布氏菌抗体辣根过氧化物酶结合物及其冻干制品基础上 ,建立检测布氏菌抗原的双抗体夹心酶联免疫吸附试验 (DAbS -ELISA) ,包括常规DAbS -ELISA及快速DAbS -ELISA。用建立的两种DAbS -ELISA法对布氏菌菌体抗原及可溶性抗原进行有关其特异性及敏感性对比观察 ;同时对模拟的这些抗原污染的饮用水标本进行对比观察。结果　常规DAbS -ELISA检测布氏菌 5 44A、16M、13 3OS及 10 4M的菌体抗原以及布氏菌 16M可溶性抗原的敏感性分别为 0 .2 2万菌 /ml、12 0万菌 /ml、190万菌 /ml、2万菌 /ml和 0 .0 12 μg/ml ;用快速DAbS -ELISA法的敏感性分别为 2 2万菌 /ml、12 0万菌 /ml、190 0万菌 /ml、2 0万菌 /ml和 1.2 μg/ml;同时用这两种DAbS -ELISA检测上述抗原污染的饮用水标本其敏感性分别为 0 .2 2万菌 /ml、190 0万菌 /ml、2 0万菌 /ml和 0 .0 12 μg/ml ;以及 2 2万菌 /ml、12 0万菌 /ml、190 0万菌 /ml、2 0 0万菌 /ml和 1.2 μg/ml。而且这两种DAbS -ELISA法检测不含布氏菌抗原的所有对照均为阴性反应。结论　双抗体夹心酶联免疫吸附试验是检测布氏菌抗原的特异、敏感、快速和适用的试验方法。Objectice In order to develop detection technique that not only specific,sensible but rapid for Brucella antigens detection.Methods A double antibodies sandwich Enzymelinked Immunosorbent Assay(DAbS-ELISA) have been developed under the basis of higher titer and freeze dried production conjugate of Brucella antibodies with horseradish peroxidase.It include DAbS-ELISA and rapid DAbS-ELISA.The specificity and sensitivety for somatic and soluble antigens from Brucella were detected by routine and rapid DAbS-ELISA.In the same time,as above antigens samples in potable water were detected too by both DAbS-ELISA.Results Of the routine DAbS-ELISAand rapid DAbS-ELISA,Sensitivety for somatic antigens from Brucella 544A,16M,133OS,104m and soluble antigen from Brucella 16M were 0.22,120,190,2(ten thousand thallus/ml)and 0.012μg/ml mycoprptein as were as 22,120,1900,20(ten thousand thallus/ml)and 1.2μg/ml mycoprotein respectively.Of the both DAbS-ELISA,sinsiticety for above antigens samples in potable water were 0.22,120,190,20(ten thousand thallus/ml) and 0.012μg/ml mycoprotein as well as 22,120,1900,200(ten thousand thallus/ml)and 1.2 μg/ml mycoprotein repectively.Above all things,Samples of no-antigens were all of negative.Conclusions The DAbS-ELISA for Brucella antigens detection were specific,sensible,rapid and practicable technique.

关 键 词：布氏菌 可溶性抗原 DA 双抗体夹心酶联免疫吸附试验 ELISA法 敏感性 对比观察 ELISA检测 菌体抗原 高滴度 

分 类 号：R378.5[医药卫生—病原生物学] R446[医药卫生—基础医学]