Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, Sichuan, China.
J Virol Methods. 2013 Apr;189(1):105-9. doi: 10.1016/j.jviromet.2013.01.007. Epub 2013 Jan 31.
In this study, a recombinant fusion antigen of duck enteritis virus (DEV) UL16 protein was expressed in Escherichia coli Rossetta (DE3). This target protein was used as a coating antigen to establish an indirect ELISA for detecting anti-DEV antibodies in serum samples from ducks. In the optimal method for the UL16-ELISA, the fusion protein was coated at 1.25μg/ml and duck serum samples were diluted at 1:160. The endpoint cut-off value of this assay was 0.598. The inter-assay and intra-assay coefficients of variation (CVs) were both lower than 10%. There was no cross-reaction with duck positive sera of either DHBV, DHV, RA, E. coli, Salmonella anatum, H5N1 or DSHDV. The assay was applied successfully to examine the suspected duck serum samples and showed 95.5% (73/76) identity with the serum neutralization test (SNT). The results showed that recombinant DEV UL16 protein could be used as a coating antigen and the developed UL16-ELISA approach was rapid, specific, sensitive and repetitive.
在这项研究中，鸭肠炎病毒（DEV）UL16 蛋白的重组融合抗原在大肠杆菌 Rosetta（DE3）中表达。该靶蛋白被用作包被抗原，以建立间接 ELISA 来检测血清样本中的抗-DEV 抗体。在 UL16-ELISA 的最佳方法中，融合蛋白以 1.25μg/ml 包被，鸭血清样本以 1:160 稀释。该测定的终点截断值为 0.598。该测定的批内和批间变异系数（CV）均低于 10%。与鸭乙型肝炎病毒（DHBV）、鸭肝炎病毒（DHV）、鸭呼肠孤病毒（RA）、大肠杆菌、沙门氏菌、H5N1 或鸭坦布苏病毒（DSHDV）的阳性血清均无交叉反应。该测定成功地应用于检测疑似鸭血清样本，与血清中和试验（SNT）的符合率为 95.5%（73/76）。结果表明，重组 DEV UL16 蛋白可用作包被抗原，所建立的 UL16-ELISA 方法快速、特异、敏感、重复性好。
