Nazerian K, Lee L F, Payne W S
U.S. Department of Agriculture, Regional Poultry Research Laboratory, East Lansing, Michigan 48823.
Avian Dis. 1990 Apr-Jun;34(2):425-32.
A highly sensitive and specific double-antibody enzyme-linked immunosorbent assay (ELISA) is described for the detection of antigen and antibody of turkey hemorrhagic enteritis virus (HEV). The assay utilizes a virus-neutralizing monoclonal antibody (MAb) to capture the antigen and turkey antiserum against HEV as the second antibody. Microtiter plates were first coated with a dilution of 1:3000 of the MAb (300 ng immunoglobulin/well) and are used for detection of both antigen and antibody. For antibody detection, MAb-coated plates were treated with an appropriate dilution of a cell-culture-propagated HEV antigen and then reacted with the test turkey serum. For detection of HEV antigen, MAb-coated plates were treated with appropriate dilutions of test antigens and then reacted with purified anti-HEV turkey immunoglobulins. The assay for HEV antibody detection was more sensitive and specific than previously described single-antibody ELISAs. Using the double-antibody ELISA, it was found that the spleen of HEV-infected turkeys harbors very high levels of antigen. Traces of HEV antigen are present in some other organs. Infectivity assay for HEV is found to be about two orders of magnitude more sensitive than the ELISA for detection of virus.
描述了一种用于检测火鸡出血性肠炎病毒（HEV）抗原和抗体的高灵敏度和特异性的双抗体酶联免疫吸附测定（ELISA）。该测定利用一种病毒中和单克隆抗体（MAb）捕获抗原，并使用针对HEV的火鸡抗血清作为第二抗体。微量滴定板首先用1:3000稀释的MAb（300 ng免疫球蛋白/孔）包被，用于抗原和抗体的检测。对于抗体检测，用适当稀释的细胞培养增殖的HEV抗原处理包被有MAb的板，然后与测试火鸡血清反应。对于HEV抗原的检测，用适当稀释的测试抗原处理包被有MAb的板，然后与纯化的抗HEV火鸡免疫球蛋白反应。HEV抗体检测测定比先前描述的单抗体ELISA更灵敏和特异。使用双抗体ELISA发现，感染HEV的火鸡脾脏中含有非常高水平的抗原。在其他一些器官中存在痕量的HEV抗原。发现HEV的感染性测定比ELISA检测病毒的灵敏度高约两个数量级。
