摘要:
利用PRV单克隆抗体-辣根过氧化物酶结合物,建立了一种敏感性高、性异特强、快速、简便的PRV抗原测定法:固相微球直接ELISA(D-ELISA)。在D-ELISA测定系统中,抗体酶结合物P3-HRP和P10-HRP的工作浓度分别为2.61μg/ml和2.85μg/ml抗体蛋白。在工作浓度条件下,D-ELISA系统可检出5.63μg/ml PRV粗提物。利用该系统测定小白鼠病理组织中的PRV抗原结果表明:同时测定额下腺耳下腺混合物、肝、脾、肺、肾等五种样本,PRV的阳性检出率可达98.0%。其中额下腺耳下腺混合物阳性率最高(87.0%),并且与阴性对照有明显的肉眼可辨的颜色差异。而其它各脏器在各批试验中,差异较大。肝的阳性率为26.3～61.3%;脾和肺均为51.8～71.4%;肾为28.6～95.0%。
Abstract:
Direct ELISA (D-ELISA) with polystyrene balls has been established to dectect PRV antigen. In the D-ELISA, the working concentrations of the conjugates, P3-HRP and P10-HRP, were 2.61 and 2.85μg /ml of Ig, respectively. With the two conjugates and their mixture, 5.63μg/ml of PRV protein (crude extract), or even lower, could be detected by the D-ELISA. The PRV antigen in the mice infected with PRV was checked by the D-ELISA with mixture, and the results indicated that positive rate of PRV antigen was 98.0% by measuring the five kinds of samples: liver, lung, spleen, kidney and submaxillary gland-subaural gland mixture. The two gland mixture had the highest positive rate (87.0%), in average and then both of lung and spleen had similla rates, 51.8-71.4%, liver 26.3-61.3%, and kidney 28.6-95.0%. The positive reaction of the two gland mixture was singnificantly different from the negative reaction of the control samples, and could be distinguished with the naked eyes. These results showed that the D-ELISA with polystyrene balls would be a highly sensitive, specific, rapid and simple technique for the clinical diagnosis of pseudorabies.
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