【摘要】 目的:建立一种应用量子点荧光标记技术检测脐血中有核红细胞γ血红蛋白表达的新方法。方法:在EDC偶联剂的作用下,将水溶性CdTe量子点与抗γ血红蛋白单克隆抗体进行连接;荧光光谱分析检测量子点偶连抗体前后的发射光谱;用免疫细胞化学法、FITC间接免疫荧光法和QDs-抗γ血红蛋白单抗荧光探针直接免疫荧光法检测脐血中有核红细胞的γ血红蛋白的表达情况。结果:QDs-抗γ血红蛋白单抗荧光探针仍然具有优良的特性,如发射光谱窄和荧光信号强;与其它两种方法相比,量子点标记法具有特异性高、荧光度强、背景清晰等优点,且量子点持续激发30min后无荧光淬灭发生。结论:偶连抗γ血红蛋白单抗的量子点荧光探针能特异性识别脐血中有核红细胞,为无痛性产前诊断奠定基础。更多还原

【Abstract】 Objective: To establish a new method of detecting γ-hemoglobin expression in nucleated red blood cells of umbilical cord blood using quantum dot fluorescence labeling. Methods: Water soluble CdTe quantum dot was conjugated to anti - γ - hemoglobin monoclonal antibody through EDC coupling reagents. The emission spectra before and after quantum dot coupled to antibody were detected through fluorescent spectroscopy. The expression of γ - hemoglobin in nucleated red blood cells of umbilical cord blood was observed by immunocytochemistry,indirect immunofluorescence using FITC and direct immunofluorescence using quantum dot - antibody fluorescent probe respectively. Results: The quantum dot - antibody fluorescent probe had still good properties such as narrow emission spectra and high florescent intensity. Compared with other two methods,CdTe quantum dot had higher brightness,better specificity and clearer background. There was no obvious photobleaching with exciting light continuous illumination for 30 minutes. Conclusion: CdTe quantum dot-antibody fluorescent probe can recognize γ - hemoglobin in nucleated red blood cells of umbilical cord blood specifically,which provides a basis for non-invasive prenatal diagnosis.更多还原