摘要
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５８份阳性，斑点杂交检出阳性份数为６２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。
Objective To develop a simple and specific assay for detection of humoral immune response in duck hepatitis B virus (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. Conclusions 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.
出处
《中华肝脏病杂志》
CAS
CSCD
2001年第1期13-15,共3页
Chinese Journal of Hepatology
