摘要
[目的]建立基于鸭胚原代肝细胞的抗乙肝药物筛选及评价模型。[方法]采用PCR技术鉴定鸭乙肝病毒(DHBV)感染鸭胚,分离感染鸭胚肝细胞原代培养,运用建立的荧光定量PCR技术检测鸭胚肝细胞DHBV-DNA复制水平,从而建立基于鸭胚原代肝细胞的抗乙肝药物评价模型。[结果]选择18～22日龄鸭胚建立了优化的鸭胚肝细胞原代培养方法和鸭胚肝细胞中DHBV-DNA水平的荧光定量PCR检测技术。[结论]该研究建立了基于鸭胚肝细胞的抗乙肝药物评价模型,经实践证实该模型具有特异、灵敏、重复性好和操作简便的优点,适用于抗乙肝化合物的筛选与评价。
[ Objective ] To establish the model for anti-hepatitis B virus （HBV） drug evaluation in duck fetal hepatocytes. [ Method ] The ordi- nary PCR assay was employed to confirm the embryonated, un-hatched, duck eggs layed by the duck hepatitis B virus （DHBV）-infected ducks. Duck fetal hepatocytes were isolated and primary cultured. The SYBR green dye-based fluorescence quantitative PCR assay was established for detection of DHBV-DNA in primary duck fetal hepatocytes. [ Result ] Duck fetal hepatocytes were isolated from 18 -22-day-old duck embryo and their optimized parameters of isolation and culture were determined. The SYBR green dye-based quantitative PCR assay for detection of DHBV- DNA was established in primary duck fetal hepatocytes. [ Conclusion] The study established a model to evaluate anti-HBV drug by the SYBR green dye-based fluorescence quantitative PCR assay for detection of DHBV-DNA in primary duck fetal hepatoeytes. Furthermore, the model was demonstrated to be a sample, specific, sensitive and repeatable assay for screening and evaluation of anti-HBV drugs.
出处
《安徽农业科学》
CAS
2013年第14期6276-6279,共4页
Journal of Anhui Agricultural Sciences
基金
教育部科技研究重点项目(212073)
科技援疆计划项目(201191260)
关键词
乙肝模型
鸭乙肝病毒
鸭胚肝细胞
原代培养
荧光定量PCR检测
Hepatitis B model
Duck hepatitis B virus
Duck fetal hepatocytes
Primary culture
Fluorescence quantitative PCR assay
