光滑型布鲁杆菌抗体竞争ELISA检测方法的建立

用提纯的布鲁杆菌疫苗株M5-90光滑型脂多糖(S-LPS)作为包被抗原,以针对S-LPS C表位的单抗16C5为竞争抗体,建立了检测光滑型布鲁杆菌血清抗体的cELISA方法.通过对临床507份牛血清样本进行检测,结果经统计分析表明,cELISA与RBT和IDEXX公司的iELISA试剂盒符合率分别为84.7%和91.2%.其中142份样品与Svanova公司的cELISA试剂盒和黑龙江省动物监察所提供的iELISA试剂盒的符合率分别为97.2%和94.4%.κ统计显示,该cELISA方法与RBT、IDEXX公司的iELISA试剂盒、Svanova公司的cELISA试剂盒和黑龙江省动物监察所提供的iELISA试剂盒的κ值均大于0.5,符合率很高.该cELISA方法为布鲁菌病的检测提供了一种有效的血清学方法,并且能够有效地排除常见的革兰阴性菌引起的假阳性结果.

Development of a competitive ELISA for detection of antibodies against smooth Brucella

A competitive enzyme-linked immunosorbent assay(cELISA) for detecting serum antibodies against smooth Brucella was developed using Brucella S-LPS C epitope McAbs 16C5 as competitive anti-body and using the purified S-LPS of Brucella vaccine strain M5-90 as coating antigen. Serum samples from cattle (n= 507) were tested by cELISA, the rose Bengal plate test(RBT) and IDEXX indirect enzyme-linked immunosorbent assay (iELISA) Kits, respectively. The coincidence between cELISA and RBT or IDEXX iELISA Kit was 84. 7% or 91. 2% respectively. 142 out of the 507 serum samples were tested by cELISA,Svanova cELISA and iELISA kit provided by Heilongjiang Animal Inspection Institute, and the results showed that the coincidence between cELISA and iELISA or Svanova cELISA Kit was 94. 4% or 97. 2%,respectively. The result by cELISA was in good agreement (κ>0. 5) with that by other method. It was concluded that cELISA with good reliability could provide a valuable tool for the serological diagnosis of brucellosis and could eliminate false-positive serological reactions caused by Yersinia enterocolitica O:9, Escherichia coli O:157 and so on.