建立双抗原夹心酶免疫试验对人畜布鲁氏菌抗体检测的研究  

Development of double antigen sandwich enzyme immunoassay for the detection of specific antibodies in humans and animals

作　　者：王显军[1] 李兰玉 赵鸿雁[2] 王勤忠[1] 冯开军[1] 李忠[1] 寇增强[1] 丁淑军[1] 张晓梅[1] 鲁齐发 

机构地区：[1]山东省疾病预防控制中心,济南250014 [2]中国疾病预防控制中心传染病预防控制所

出　　处：《中国人兽共患病学报》2007年第3期262-265,共4页Chinese Journal of Zoonoses

基　　金：山东省医药卫生科技项目课题(No.2001CA1DDB1)

摘　　要：目的为人畜布鲁氏菌病(布病)的血清学诊断、监测及流行病学调查提供特异、敏感、快速的试验方法。方法在制备高滴度的布鲁氏菌(布氏菌)抗原及其抗原辣根过氧化物酶结合物基础上。建立了检测人畜布氏菌抗体的双抗原夹心酶免疫试验(DAgS-EIA),包括常规双抗原夹心酶联免疫吸附试验(DAgS-ELISA)、双抗原夹心酶免疫斑点试验(DAgS-DIEA)以及快速双抗原夹心ELISA(快速DAgS-ELISA),并对影响本试验的主要因素因素进行了试验。结果制备了高滴度的布氏菌抗原及其抗原酶结合物和冻干制品。并在此基础上,建立了常规DAgS-ELISA及DAgS-DIEA以及快速DAgS-ELISA检测人畜布氏菌抗体方法。经对115份布病患者血清检测结果,阳性率以DAgS-ELISA为最高(61.7%),其余依次为RBPT(58.1%)I、-ELISA(55.6%)、DAgS-DIEA(53.7%)、及SAT(44.2%)且用DAgS-EIA检测布氏菌感染羊、牛、猪及实验动物家兔、豚鼠和小鼠均为强阳性反应,而对正常人、羊、牛、猪及实验动物血清均为阴性反应。结论双抗原夹心酶免疫试验检测人畜血清布氏菌抗体,不仅特异、敏感、快速、简便而且仅需制备单一的布氏菌抗原酶结合物就可对人及各类动物血清布氏菌抗体进行检测。In order to develop a specific, sensitive ,rapid and practical method for serodiagnosis, surveillance and epidemiological surveys of brucelosis in human and animals, the double antigen sandwich enzyme immunoassays（DagS-EIA） inclucling routine double antigen sandwich enzyme linked absorbent assay （DagS-ELISA）, double antigen sandwich enzyme immunospot assay, （DAgS-DIEA） and rapid double antigen sandwich enzyme immunoassay （rapid DagS-ELISA） had been developed on the basis in the preparation of high-titered Brucella antigen and its conjugate of Brucella antigen with horseradish peroxidase. In addition, the main factors influencing the result of DagS-EIA had been evaluated. In the present study, the high-tittered Brucella antigens and the freezing dried conjugate of Burucella antigen and hoseradish peroxidase had been prepared, and on the basis of these preparations, the DagS-EIA was established for the detection of the Brucella antibodies in humans and animals. Upon the detection of Burucella antibodies in 115 serum samples of patients with brucellosis, it was found that positive rates of antibody detection with DagS-ELISA, RBPT, IELISA, DagS-DIEA and SAT were 61.7%, 58.1%, 55.6%, 53.7% and 44. 2% respectively, When these methods were applied for the detection of Brucella antibodies in sheep, cattle and swine as well as experimental animals such as rabbit, guinea pig and mice infected with Brucella, all showed strong positive reactions. However, no positive reaction could be detected in sera of normal human, sheep, cattle or swine. It is concluded that the DagS-EIA is not only specific, sensitive, rapid and practicable, but also may be used to detect various Brucella antibodies in human and animals by the preparation of one conjugate for Brucella antigen with horseradish peroxidase.

关 键 词：布鲁氏菌 酶免疫试验 抗原 抗体 

分 类 号：R378.5[医药卫生—病原生物学]