中文摘要：

目的建立检测博尔纳病病毒（BDV）抗原的双抗体夹心ELISA法,为简单、快速的临床应用奠定基础。方法根据双抗夹心ELISA法基本步骤,分别对BDV p24小鼠单克隆抗体、BDV p24兔多克隆抗体和酶标抗体作多梯度稀释,以确定最佳实验条件,并对其精密度、灵敏度、稳定性、准确性进行初步评价,同期收集病毒性脑炎（VE）患者脑脊液（CSF）44例和神经系统非炎性疾病患者CSF 44例,通过建立的双抗夹心ELISA法检测临床样本中是否存在BDV抗原。结果 BDV p24小鼠单克隆抗体的最佳稀释度为1∶160,BDV p24兔多克隆抗体的最佳稀释度为1∶2 500,酶标抗体的最佳稀释度为1∶5 000,该法的检测灵敏度为170 ng/ml,精密度批内变异〈10%,批间变异〈15%,同时具有良好的稳定性与准确性。另外用该法检测临床标本其中实验组有3例患者检出BDV抗原阳性,阴性对照组均未检出阳性病例。结论成功建立检测BDV抗原的双抗体夹心ELISA法。

英文摘要：

In this study, we aimed to establish a double-antibody sandwich ELISA for detecting the antigen of Borna disease virus （BDV）. Firstly, we explored important experimental parameters （the dilution of BDV p24 mouse monoclonal antibody, the dilution of BDV p24 rabbit polyclonal antibody, and the dilution of the enzyme-labeled antibody） by the double-antibody sandwich ELISA specific steps. Then, we evaluated precision, sensitivity, stability and accuracy of established assay. At last, we collected 44 cases of cerebrospinal fluid （CSF） from viral encephalitis （VE） patients and 44 cases from non-inflammatory diseases of the nervous system to detect the existence of BDV antigen with the established double-antibody sandwich ELISA. Our result indicated the optimal dilution of BDV p24 mouse monoclonal antibody was 1：160, the optimal dilution of BDV p24 rabbit polyclonal antibody was 1：2 500, and the optimal dilution of enzyme-labeled antibody was 1：5 000. Besides, the sensitivity of double-antibody sandwich ELISA was 170 ng/ml, precision batch variation was less than 10%, and inter-assay variation was less than 15%. Meanwhile, there were three cases to be detected with the BDV antigen in the experimental group, while cases in negative control group can not be detected with BDV antigen. In summary, the study successfully established a double-antibody sandwich ELISA for detecting BDV antigen, which posses better stability and accuracy.