鸭乙型肝炎病毒表面抗原的纯化及应用 |
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引用本文: | 唐霓,黄爱龙,郭树华,张定凤.鸭乙型肝炎病毒表面抗原的纯化及应用[J].重庆医科大学学报,2001,26(1):14-16. |
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作者姓名: | 唐霓 黄爱龙 郭树华 张定凤 |
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作者单位: | 重庆医科大学病毒性肝炎研究所 |
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摘 要: | 目的:从鸭乙型肝炎病毒(DHBV)感染鸭血清中纯化表面抗原(DHBsAg),初步应用于病毒感染后复制的研究。方法:用蔗糖密度梯度离心纯化病毒表面抗原,经Bradford法定量血清中该蛋白含量;建立DHBsAgELISA检测方法,并与DHBVDNA斑点杂交相比较。结果:纯化DHBsAg经ELISA检测OD490nm≥1.0,Western杂交显示主要肽分子量为17KDa和35KDa:Bradford法定量血清蛋白含量为2.85-5.10ug/ml,DHBsAgELISA检测100份血清标本阳性数为84例,斑点杂交检测阳性数为88例,两者吻合率为94%。结论:DHBsAg的制备及ELISA方法的建立,为进一步研究病毒感染后复制及体内免疫应答状况奠定了基础。
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关 键 词: | 鸭乙型肝炎病毒表面抗原 Western杂交 纯化 DHBV |
文章编号: | 0253-3626(2001)01-0014-03 |
修稿时间: | 2000年1月11日 |
Preparation and identification of duck hepatitis B virus (DHBV) surface antigen from infected duck serum |
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Abstract: | Objective: Preparation of DHBsAg from DHBV infected ducks and its application for the study of virus replication. Methods: The positive serum were layered over a step gradient of 20% and 66% sucrose,DHBsAg were harvested at the interface between 20% and 66% sucrose, and then analyzed by Bradford method.Meanwhile, DHBsAg ELISA and DHBV DNA dot blot hybridization assayed 100 serum samples. Results:Purified antigen were detected OD490nm≥1.0 by DHBsAg ELISA assay and quantified 2.85~5.10μg/ml by Bradford method. Both 35KDa and 17KDa pres/s bands were present in DHBV-infected serum. DHBsAg assayed 84 positive in 100 serum samples, while DHBV DNA dot blot hybridization detected 88 positive, showing good accordance. Conclusion: Preparation of DHBsAg from DHBV-infected ducks and application of DHBsAg ELISA assay will be helpful to the study of kinetics of DHBV and immune response of ducks to the virus. |
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