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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 a gold standard Fn-EIIIA monoclonal antibody test strip that can be used to estimate the probability of forensic pathology injury. Methods Two sensitive and specific fragment-specific monoclonal antibodies to Fn-EIIIA were selected. The titer was determined by ELISA. The colloidal gold was labeled with gold nanoparticles and their effects were determined by percolation and chromatography. The target protein was extracted from the positive control Preliminary evaluation, and rat skin damage model validation. Results Both monoclonal antibodies could be used to detect Fn-EIIIA peptide by gold standard method, and IST9 with Fn polyclonal antibody had the best effect. The test strip in normal skin tissue samples did not show any detectable color after 3h of injury, With the extension of injury time, staining gradually deepened. Conclusion The test strip reliability, repeatability and maneuverability are good, is expected to foresee a wide range of applications in forensic injury time.