National Avian Influenza Reference Laboratory, Animal Influenza Laboratory of the Ministry of Agriculture, Harbin Veterinary Research Institute, CAAS, Harbin 150001, China.
J Virol Methods. 2010 Oct;169(1):66-9. doi: 10.1016/j.jviromet.2010.06.018. Epub 2010 Jul 4.
The VP1-encoding gene of the duck hepatitis type 1 virus (DHV-1) HP-1 strain was cloned and expressed in Escherichiacoli. The open reading frame (ORF) of VP1 comprised 714 bp and encoded 238 amino acids, with a predicated molecular mass of 26.5 kDa. The expressed VP1 fusion protein in E. coli was detected by Western blotting with duck anti-DHV-1 polyclonal serum. A VP1-ELISA using the expressed VP1 protein as a coating antigen for the detection of antibodies to DHV-1 in ducks was developed. In comparison with the virus neutralization test, the specificity and sensitivity of the VP1-ELISA was 92.5% and 96.7%. Comparative analysis between Western blots and the VP1-ELISA showed that the concordance between the two methods was 86%. The VP1-ELISA did not react with the anti-sera to other duck viral pathogens, implying that this protein is specific for the recognition of duck anti-DHV-1 antibodies. Taken together, the VP1-ELISA is a highly sensitive and specific test that could be used for screening for DHV-1 infection and monitoring antibody titres against DHV-1.
鸭肝炎 1 型病毒（DHV-1）HP-1 株的 VP1 编码基因在大肠杆菌中被克隆和表达。VP1 的开放阅读框（ORF）由 714bp 组成，编码 238 个氨基酸，预测分子量为 26.5kDa。用鸭抗 DHV-1 多克隆血清通过 Western blot 检测到大肠杆菌中表达的 VP1 融合蛋白。建立了一种使用表达的 VP1 蛋白作为包被抗原检测鸭 DHV-1 抗体的 VP1-ELISA。与病毒中和试验相比，VP1-ELISA 的特异性和灵敏度分别为 92.5%和 96.7%。Western blot 和 VP1-ELISA 的比较分析表明，两种方法的一致性为 86%。VP1-ELISA 与针对其他鸭病毒性病原体的抗血清不反应，表明该蛋白特异性识别鸭抗 DHV-1 抗体。综上所述，VP1-ELISA 是一种高度敏感和特异的检测方法，可用于筛查 DHV-1 感染和监测针对 DHV-1 的抗体滴度。
