Recombinant DNA Laboratory, Indian Veterinary Research Institute, Izatnagar, Bareilly, Uttar Pradesh 243122, India.
J Virol Methods. 2012 Nov;185(2):234-8. doi: 10.1016/j.jviromet.2012.06.027. Epub 2012 Jul 4.
A recombinant UL30 antigen-based single serum dilution enzyme linked immunosorbent assay (ELISA) was developed to measure specific antibody in the sera of ducks against duck enteritis virus (DEV). The partial UL30 gene of DEV was cloned, expressed, purified and tested for its diagnostic use by designing a single serum dilution enzyme linked immuno-sorbent assay (ELISA). A total of 226 duck sera samples were tested using the assay. A linear relationship was found between the predicted antibody titres at a single working dilution of 1:100 and the corresponding serum titres observed as determined by the standard serial dilution method. Regression analysis was used to determine a standard curve from which an equation was derived which demonstrated this correlation. The equation was then used to convert the corrected absorbance readings of the single working dilution directly into the predicted ELISA antibody titres. The assay proved to be specific, sensitive and accurate as compared to the virus neutralization test with a specificity, sensitivity and accuracy being 96%, 95% and 95% respectively.
建立了一种基于 UL30 重组抗原的单份血清稀释酶联免疫吸附试验（ELISA），用于检测鸭血清中针对鸭肠炎病毒（DEV）的特异性抗体。通过设计单份血清稀释 ELISA，克隆、表达、纯化了 DEV 的部分 UL30 基因，并对其诊断用途进行了测试。使用该检测方法对 226 份鸭血清样本进行了检测。在 1:100 的单一工作稀释度下，预测的抗体效价与标准系列稀释法观察到的相应血清效价之间存在线性关系。通过回归分析，从标准曲线上得出一个方程，证明了这种相关性。然后，该方程用于将单份工作稀释的校正吸光度读数直接转换为预测的 ELISA 抗体效价。与病毒中和试验相比，该检测方法具有特异性、敏感性和准确性，特异性、敏感性和准确性分别为 96%、95%和 95%。
