应用ELISA快速检测牛羊副结核病

Rapid Detection of Paratuberculosis in Cattle and Sheep by ELISA

在96孔微板中包被抗原,加入被检血清,经温育后,抗体特异性地与抗原相结合,形成抗原-抗体结合物。洗涤去除未结合的物质,与加入的酶结合物(HRPO)相结合。再洗去未结合的酶结合物,加入底物后显色,加入终止液终止反应。用装有630nm(或620nm、650nm)滤光片的酶标仪进行检测,通过计算S／P值,判定血清中副结核抗体阴、阳性。此方法具有快速、灵敏、准确的特点。

Test sera were added to antigen - coated microplate in an ELISA for dectection of paratuberculosis. After incubation and washings, HRPO was added.Results were read after colour development and stopping of recation stopped.S/ P<0.25 was considered as negative and S/ P≥0.25 as positive for paratuboculosis antibodies.Test with positive and negalive controls and 10 serum samples showed that the assay was rapid, sensitive and accurate. Absorption of serum with Mycobacterium phlei prior to the actual test would improve the specificity.