Fn-EIIIA金标单克隆抗体试纸条的试制和初步应用  

Preliminary Application on Test Paper of Gold-labelled Antibody against Fibronectin EⅢA Splicing Variant

作　　者：刘宁国[1] 陈忆九[1] 卢仁泉 黄晓华[1] 张建华[1] 

机构地区：[1]司法部司法部司法鉴定科学技术研究所,上海200063 [2]上海本草生物医学工程研究所,上海200040

出　　处：《法医学杂志》2006年第2期81-83,共3页Journal of Forensic Medicine

基　　金：上海市自然科学基金项目(05214118)

摘　　要：目的开发可用于法医病理学损伤时间推断临案的金标Fn-EIIIA单克隆抗体试纸条。方法选取两种敏感、特异的片段特异性Fn-EIIIA单克隆抗体,ELISA方法进行效价观察,标记胶体金后分别采用渗滤法及层析法检测其效果,抽提阳性对照中目标蛋白进行初步测评,并以大鼠皮肤损伤模型进行验证。结果两种单克隆抗体均可用于Fn-EIIIA肽段的金标法检测,以IST9搭配Fn多抗效果最佳;正常皮肤组织样本中试纸条检测线不显色,损伤3h后显色,随损伤时间延长,染色逐渐加深。结论该试纸条可靠性、可重复性以及可操作性均佳,可望在法医学损伤时间推断有广阔的应用。Objective To develop the test Paper of gold-labelled antibody against fibronectin EⅢA splicing variant, which can be available in forensic wound interval estimation. Methods Two sensitive antibodies were compared with enzyme link immunoabsorband assay （ELISA）. After colloid gold labeled, the effects of the two antibodies were tested by methods of Dot immunogold filtration assay and gold immunochromatography assay, respectively. The test paper was finally appraised by applied in experimental skin injury in rats. Results On the test paper, detected line appeared in three hours wound age group,anu then the darkness ol positive staining became darker with injury time prolonging, while control normal skin cannot found to be positive staining. Conclusion The gold-labeled test paper can be useful in estimation of wounding interval in forensic science.

关 键 词：纤维连接蛋白 可变剪接 额外结构域 EⅢA 损伤时间推断 胶体金 

分 类 号：R392-33[医药卫生—免疫学] R446.12[医药卫生—基础医学]