鸭乙型肝炎病毒体液免疫血清学指标的系统建立与应用

摘要

Objective To develop a simple and specific assay for detection of humoral immune response in DHBV infected ducks. Methods Eighty serum samples were detected by ELISA for DHBsAg with prepared anti-preS 1H1 ascitic fluid and by PCR or dot blot hybridization for DHBV DNA. Thirty-two serum samples from experimentally infected 1-day-old ducks were assayed by ELISA for DHBcAb and DHBsAb. Results Of 66 PCR positive samples, 58 were positive for DHBsAg and 62 were positive for DHBV dot blot hybridization. Sensitivities of the two methods were 87.9% and 93.9%, respectively. Anti-DHBc was developed in 2/8 infected ducks at day 21 but negative after day 28. Anti-DHBs antibodies were negative throughout the infectious period. Conclusion The application of DHBV infection and serological assay will be helpful to the study of kinetics of DHBV and humoral immune response of ducks against the virus.%目的　建立简便、特异的鸭乙型肝炎病毒（DHBV）感染及免疫血清学检测系统。方法制备、纯化检测所需的抗DHBV前S区单克隆抗体腹水、DHBsAg和rDHBcAg，对随机筛选的80份感染血清及实验感染1日龄重庆麻鸭系列血清分别进行DHBsAg和抗-DHBc、抗-DHBs检测。结果 PCR方法检测出阳性66份，阴性14份。选用PCR阳性的66份标本，经DHBsAg ELISA检测出62份阳性，斑点杂交检出阳性份数为58份。与DHBV PCR相比较，灵敏度分别为87.9%和 93.9%，而8只实验感染鸭仅2只在感染后第3周血清抗-DHBc阳性，抗体效价为1:10，至第4周测定已为阴性，抗-DHBs则在所有标本均为阴性。结论 DHBV感染及免疫血清学酶联免疫方法的建立，为进一步研究DHBV感染后复制规律及体内免疫应答状况奠定了基础。