猪O型口蹄疫病毒抗体化学发光酶联免疫检测方法的建立

Establishment of chemiluminescent enzyme immunoassay for detecting antibodies against foot-and-mouth disease virus serotype O in swine

摘要:

表达并纯化猪O型口蹄疫病毒 (FMDV) VP1重组蛋白作为检测抗原，建立了一种快速检测猪O型口蹄疫病毒抗体的化学发光酶联免疫 (CLEIA) 检测方法。建立的VP1-CLEIA方法特异性为100%，板内变异系数在1.10%–6.70%之间，板间变异系数在0.66%–4.80%之间，具有较好的特异性和重复性，且灵敏度高于ELISA方法。通过对山东、辽宁、河北地区采集的250份临床血清的检测表明，该方法与间接ELISA试剂盒的符合率为93.50%，与液相阻断ELISA试剂盒的符合率为94.00%，表明本次建立的VP1-CLEIA检测方法可以用于猪O型FMDV感染或疫苗免疫后抗体水平检测。

Abstract:

Recombinant structural protein VP1 of foot-and-mouth disease virus serotype O was expressed in Escherichia coli and then purified using Nickel affinity chromatography. A chemiluminescent enzyme immunoassay (CLEIA) method was established using the purified recombinant protein as coating antigen to detect antibody of foot-and-mouth disease virus serotype O in swine. The specificity of VP1-CLEIA method is 100%. The coefficients of variation in the plate and between plates are 1.10%–6.70% and 0.66%–4.80%, respectively. Comparing with the commercial indirect ELISA kit or liquid phase block ELISA kit, the calculated coincidence rate is 93.50% or 94.00%. The high specificity and stability suggested this detection method can be used to monitor the antibody level of foot-and-mouth disease virus serotype O in swine.