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目的研究免疫应答和耐受HBeAg阳性慢性乙型肝炎患者红细胞补体受体Ⅰ型分子(CR1)基因点突变与HBV DNA含量变化的关系,探讨其临床意义。方法选择肝脏功能正常、血清HBV DNA大于106IU/ml的114例HBeAg阳性慢性乙型肝炎患者作为免疫耐受组研究对象,选择初次发生转氨酶高于正常值2倍以上伴或不伴有肝细胞性黄疸的110例HBeAg阳性慢性乙型肝炎患者为免疫应答组研究对象。采用PCR和HindⅢ酶切技术对红细胞CR1分子基因多态性进行分类。用细胞核酸释放剂(CNR)裂解细胞,释放HBV DNA,对白细胞HBV DNA进行实时荧光PCR检测。结果免疫应答组HBeAg阳性慢性乙型肝炎患者的红细胞CR1基因点突变率明显低于免疫耐受组和正常人群(P<0.05)。在免疫应答组中,CR1基因发生点突变患者的血清和白细胞HBV DNA含量均明显高于未发生点突变者(P<0.05);在免疫耐受组中,红细胞CR1基因发生点突变和无点突变的患者其血清和白细胞HBV DNA差异均无统计学意义。结论检测HBeAg阳性慢性乙型肝炎患者红细胞CR1基因点突变对了解乙型肝炎患者清除HBV的能力、评估病情发展、判断预后具有一定的指导意义。
Objective To study the relationship between the point mutation of erythrocyte complement receptor type 1 (CR1) gene and HBV DNA content in patients with immune response and HBeAg-positive chronic hepatitis B and to explore its clinical significance. Methods 114 patients with HBeAg-positive chronic hepatitis B who had normal hepatic function and serum HBV DNA greater than 106 IU / ml were selected as the subjects of immune tolerance group. The patients with primary liver transaminase more than 2 times of the normal value were selected with or without hepatocellular 110 cases of jaundice HBeAg-positive chronic hepatitis B patients for the immune response to the study. PCR and Hind Ⅲ restriction enzyme digestion of CR1 molecular gene polymorphism of red blood cells were classified. Cells were lysed with cellular nucleic acid release (CNR), HBV DNA was released, and white blood cell HBV DNA was detected by real-time fluorescence PCR. Results The point mutation rate of erythrocyte CR1 gene in patients with HBeAg-positive chronic hepatitis B was significantly lower than that in immunocompromised and normal controls (P <0.05). In the immune response group, the levels of serum and leucocyte HBV DNA in patients with point mutations of CR1 gene were significantly higher than those without the point mutation (P <0.05). In the immune tolerance group, the CR1 gene of erythrocytes was mutated and no point Mutant patients had no significant difference in serum HBV DNA and white blood cells. Conclusion Detection of erythrocyte CR1 gene mutations in patients with HBeAg-positive chronic hepatitis B is of guiding significance in understanding the ability of hepatitis B patients to clear HBV and assessing the progression of disease and prognosis.