| 胶体金探针对HBV全基因转染滋养细胞中HBsAg的示踪研究 |
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| 引用本文： | 张磊,邵晨,邵中军,张景霞,卢娟,门可,闫永平,徐德忠. 胶体金探针对HBV全基因转染滋养细胞中HBsAg的示踪研究[J]. 疾病控制杂志, 2010, 14(2): 99-103 |
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| 作者姓名： | 张磊 邵晨 邵中军 张景霞 卢娟 门可 闫永平 徐德忠 |
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| 作者单位： | 1. 第四军医大学流行病学教研室,陕西,西安,710032
2. 第四军医大学西京医院泌尿外科,陕西,西安,710033 |
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| 基金项目： | 国家自然科学基金重点课题(30230320);;陕西省自然科学基金(2006C248) |
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| 摘 要： | 目的通过胶体金探针标记乙肝病毒（HBV）全基因转染的人胎盘滋养细胞（HPT-8-HBV），示踪乙肝表面抗原（HBsAg）在HPT-8-HBV细胞中的分布。方法通过脂质体介导的方法将HBV全基因转染HPT-8细胞，G418筛选，通过ELISA、免疫组化对转染细胞进行检测；采用胶体金标记的HBsAg单克隆抗体作为探针（简称胶体金探针）对转染细胞中的HBsAg进行标记示踪。结果成功将质粒pcDNA3—3HBV转染到HPT-8，转染后第1、2代培养上清ELISA检测显示HBsAg阳性，免疫组化检测第2代转染细胞中HBsAg阳性；通过胶体金探针标记发现HBsAg存在于转染滋养细胞中的粗面内质网、分泌泡、空泡和溶酶体内，以及细胞膜和绒毛上。结论可以从重组质粒pcDNA3—3HBV转染的滋养细胞的树脂切片中获得HBsAg的超微结构定位。
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| 关 键 词： | 肝炎表面抗原 乙型 滋养细胞 探针 转染 |
Tracer study of human placental trophoblast cell (HPT-8) transfected with pcDNA3-3HBV using anti-HBs bound colloidal-Gold probes |
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| ZHANG Lei,SHAO Chen,SHAO Zhong-jun,ZHANG Jing-xia,LU Juan,MEN Ke,YAN Yong-ping,XU De-zhong. Tracer study of human placental trophoblast cell (HPT-8) transfected with pcDNA3-3HBV using anti-HBs bound colloidal-Gold probes[J]. Chinese Journal of Disease Control and Prevention, 2010, 14(2): 99-103 |
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| Authors: | ZHANG Lei SHAO Chen SHAO Zhong-jun ZHANG Jing-xia LU Juan MEN Ke YAN Yong-ping XU De-zhong |
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| Affiliation: | 1. Department of Epidemiology, Fourth Military Medical University, Xi ＇ an 710032, China ; 2. Department of Urology, Xijing Hospital, Fourth Military Medical University, Xi＇an 710033, China) |
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| Abstract: | Objective To trace the distribution of hepatitis B surface antigen(HBsAg) in human placental trophoblast(HPT-8-HBV) cells transfected by pcDNA3-3HBV using anti-HBs bound colloidal-Gold probes.Methods HPT-8 cells were transfected with pcDNA3-3HBV and screened with antibiotic G418.The transfected cells were detected by ELISA and immunocytochemical staining.HBsAg in transfected cells were marked and traced using an anti-HBs bound colloidal gold probes,combined with transmission electron microscopy technique.Re... |
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| Keywords: | Hepatitis B surface antigen Trophoblast cell Probe Transfection |
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