Shien J H, Yin H S, Lee L H
Department of Veterinary Medicine, National Chung Hsing University, Taichung, Taiwan 403, Republic of China.
Res Vet Sci. 2000 Oct;69(2):107-12. doi: 10.1053/rvsc.2000.0414.
An enzyme-linked immunosorbent assay using the expressed protein sigma B as the coating antigen (sigma B-ELISA) for detecting antibody to avian reovirus (ARV) in chickens was developed and compared with a conventional ELISA. Both ELISA s and a serum neutralisation (SN) test were used to test the sera from experimentally vaccinated and farm chickens. The sigma B-ELISA could clearly distinguish the SN-positive and -negative sera in 38-week-old chickens. The correlation rate between SN and a sigma B-ELISA was 100 per cent (65/65), and that between SN and conventional ELISA was 84 per cent (55/65). With the sigma B-ELISA, all SN-negative sera had low absorbance values (below 0.06), and the absorbance values correlated closely with the SN titres. However, the sera which were antibody-negative by SN had various absorbance values, ranging from 0.07 to 0.39 in the conventional ELISA. Hence, the sigma B-ELISA had lower non-specific binding reactions than the conventional ELISA against sera from ARV -negative birds. Antibody against ARV could be detected by sigma B-ELISA after vaccination. Absorbance values peaked 4 weeks after vaccination at 2 weeks of age and were maintained until the birds were 27 weeks old. The results suggest that the presence of antibody against viral protein sigma B in birds may be used as a good indicator by the sigma B - ELISA for detecting immune status of a chicken flock or to detect chickens infected with ARV.
建立了一种以表达的σB蛋白为包被抗原的酶联免疫吸附测定法(σB-ELISA),用于检测鸡群中禽呼肠孤病毒(ARV)抗体,并与传统ELISA进行比较。两种ELISA方法以及血清中和(SN)试验均用于检测经实验性接种疫苗的鸡和农场鸡的血清。σB-ELISA能够清晰地区分38周龄鸡的SN阳性和阴性血清。SN与σB-ELISA之间的符合率为100%(65/65),SN与传统ELISA之间的符合率为84%(55/65)。使用σB-ELISA时,所有SN阴性血清的吸光度值均较低(低于0.06),且吸光度值与SN滴度密切相关。然而,SN抗体阴性的血清在传统ELISA中的吸光度值各不相同,范围为0.07至0.39。因此,与传统ELISA相比,σB-ELISA对ARV阴性鸡血清的非特异性结合反应更低。接种疫苗后,可通过σB-ELISA检测到抗ARV抗体。2周龄雏鸡接种疫苗后4周吸光度值达到峰值,并一直维持到鸡27周龄。结果表明,禽体内抗病毒蛋白σB抗体的存在可被σB-ELISA用作检测鸡群免疫状态或检测感染ARV鸡的良好指标。