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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。
OBJECTIVE: To establish a new method to detect the expression of γ-hemoglobin in nucleated red blood cells in cord blood by using quantum dot fluorescence labeling. METHODS: The water-soluble CdTe QDs were linked to anti-γ-hemoglobin monoclonal antibody by EDC coupling agent. The emission spectra of the coupled QDs were detected by fluorescence spectroscopy. The immunofluorescence and FITC indirect immunofluorescence Fluorescence and QDs-anti-γ-hemoglobin monoclonal antibody fluorescent probe direct immunofluorescence detection of cord blood erythrocytes γ-hemoglobin expression. Results: QDs-anti-γ-hemoglobin monoclonal antibody fluorescent probe still has excellent properties, such as narrow emission spectrum and strong fluorescence signal; compared with the other two methods, the quantum dot labeling method has the advantages of high specificity, strong fluorescence and clear background And other advantages, and no quenching occurred after 30min excitation of quantum dots. CONCLUSION: Quantum dot fluorescence probe coupled with anti-γ-hemoglobin monoclonal antibody can specifically identify nucleated erythrocytes in umbilical cord blood and lay a foundation for painless prenatal diagnosis.