鸭乙型肝炎病毒感染特异细胞介导免疫检测方法的建立

目的：建立简便、特异的鸭乙型肝炎病毒（DHBV）感染特异细胞介导免疫（CMI）检测方法。方法：分别从DHBV强阳性的血清和肝组织中纯化DHBsAg、DHBcAg,Bradford法定量后，用于急性DHBV感染重庆麻鸭CMI的检测。结果：急性感染后10d鸭外周血单个核细胞（PBMC）快速出现对DHBsAg、DHBcAg的不同程度增殖反应，5周内下降至正常水平。7份刺激细胞上清DuIFN-γ检测结果为阳性，OD540nm读数介于0．062－0．108之间，细胞因子的表达水平与PBMC增殖反应强度呈现较好的相关性。结论：DHBV感染重庆麻鸭CMI检测方法的建立，为了解感染鸭特异免疫应答状态提供了重要参考。

The development of a simple and sensitive assay for detection of cell-mediated immune response against duck hepatitis B virus

Objective To establish a simple and sensitive assay for detecion of cell mediated immune(CMI) response against duck hepatitis B virus.Methods Duck hepatitis surface and core antigen, which purified from DHBV positive serum or liver and quantitated by Brandford method respectively, were used as the specific antigen that primed CMI response in acutely infected ducks.Results Significant specific cellular response of PBMC to DHBsAg and DHBcAg were produced as early as 10 days post infection and declined to normal level in subsequent 5 weeks. Seven samples that derived from supernatant of specific antigen primed PBMC were positive for duck interferon gamma. The value of OD540nm varied from 0 062 to 0 108.The level of iduced cytokine showed a good correlation with the intensity of CMI response.Conclusion These methodologies regarding CMI response will be helpful to the study of specific immune response of duck against this virus.