直接免疫细胞化学法检测红细胞内血红蛋白及其在血红蛋白病诊断中的应用

DIRECT IMMUNOCYTOCHEMICAL DETECTION OF HEMOGLOBIN TYPES IN RED CELLS AND ITS USE IN DIAGNOSIS OF HEMOGLOBINOPATHIES

目的 :建立直接免疫细胞化学法检测红细胞内血红蛋白类型 ,以鉴别胎儿有核红细胞 ,开展从母血分离胎儿细胞进行遗传病无创性产前诊断研究。方法 :用荧光素标记的 anti- Hb A,anti- α- globin,anti- Hb E,anti- β- globin和 anti- γ- globin抗体特异性检测正常成人外周血、 Hb Bart's水肿死胎心脏血、血红蛋白 E杂合子患者和 β°地中海贫血纯合子患者外周血的细胞涂片 ,在荧光显微镜下观察阳性细胞。结果 :正常成人外周血和 Hb E杂合子患者外周血的细胞涂片在荧光显微镜下分别观察到anti- Hb A,anti- Hb E阳性细胞。Hb Bart's水肿死胎心脏血的细胞涂片 ,未见 anti- α- globin阳性细胞 ,可见 anti- γ- globin阳性细胞。β°地中海贫血纯合子患者外周血的细胞涂片 ,未见 anti- β- globin阳性细胞 ,可见 anti- γ- globin阳性细胞。结论 :直接免疫细胞化学法是一种简便有效检测红细胞内血红蛋白的方法 ,可用于鉴别从母血中分离的胎儿有核红细胞 ,开展无创性产前基因诊断研究 ,并进行生理、病理和治疗状态下特异红细胞族群的标识、分离、富集。

Objective:Establishment of detecting hemoglobin types in red blood cells with direct immunocytochemistry Methods: Cell smears of normal adult peripheral blood,heart blood of Hb Bart's hydrop fetus,peripheral blood of HbE heterozygote and β° thalassemia homozygote were specifically incubated with Fluorescein Isothiocyanate(FITC) conjugated anti HbA ,anti α globin,anti HbE and anti β globin and anti γ globin antibodies Results:Fluorescent positive red cells of anti HbA,anti HbE were observed on the blood smear of peripheral blood from normal adult and HbE heterozygote There were no fluorescent positive red cells of anti β globin and anti α globin on the blood smear of whole blood from β° thalassemia homozygote and Hb Bart's hydrop fetus,respectively Conclusion:Direct immunocytochemistry is an effective method to detect hemoglobin types in red cells With direct immunocytochemistry,the populations of red cells can be labeled,separated and enriched,traced and detected so as to achieve diagnosis,evaluation of treatment and non invasive prenatal diagnosis on hemoglobinopathies