应用MGB探针检测人乙型肝炎病毒的基因型  

Apply the MGB probe to detect HBV virus genotyping of the people

作　　者：陈泽惠 李欣钰[2] 肖湘文[2] 丁渭[2] 刘淑园[2] 陈华云[2] 

机构地区：[1]孝感市云梦县人民医院输血科,湖北孝感432500 [2]中山大学达安基因诊断中心,广东广州510665

出　　处：《分子诊断与治疗杂志》2014年第4期250-253,共4页Journal of Molecular Diagnostics and Therapy

基　　金：国家"艾滋病和病毒性肝炎等重大传染病防治"科技重大专项(2012ZX10002005)

摘　　要：目的检测血浆或血清样本中人乙型肝炎病毒HBV的基因型,建立了基于MGB探针的实时荧光定量PCR检测体系。方法在HBV基因组型特异性碱基的聚集区域设计荧光定量PCR引物及探针,为保证特征特异性及高分辨率,选择设计MGB探针,然后将PCR扩增的产物纯化后进行基因克隆,克隆成功后提取的质粒制备为阳性质控品,从而建立完整的MGB探针法的荧光定量PCR检测体系,最后对该检测体系进行方法学评价。结果成功建立了基于MGB探针的HBV荧光定量PCR的基因分型检测体系。方法学评价显示,该检测体系的灵敏度达50 IU/mL;重复性好;特异性良好,以多种血浆病毒的核酸样品为模板进行检测时结果全为阴性。结论本研究建立基于MGB探针的HBV荧光定量PCR的基因分型检测方法,可以很好地定性的检测血浆或血清样本及血液制品中的人乙型肝炎病毒HBV的基因型及含量,对于该疾病的临床治疗具有重要的意义。Objective A Real-Time PCR assay based on the MGB probe for detecting genotypes of hepatitis B virus existed in plasma or serum samples was built. Methods To ensure the specificity and high resolution, the primer and probe of Real-Time PCR of HBV was designed in the areas which genome type concentrated. To evaluate the methodology for the Real-Time PCR detection system of the MGB probe, we constructed plasmids as the positive quality products to build complete detection system. Results The detection system of HBV genotypcs of Real-Time PCR based on the MGB probe was established successfully. Methodological evaluation showed that the sensitivity was up to 50 IU/mL and repeatability is excellent. The tests of a varieties of plasma virus nucleic acid were negative to reveal high specificity. Conclusion A detection methods based on the MGB probe of HBV gcnotypes of Real-Time PCR was built to detect hepatitis B virus genotype and the content of blood, plasma or serum samples. The system is of great importance for the clinical treatment of HBV virus.

关 键 词：乙型肝炎病毒HBV MGB探针 荧光定量PCR 

分 类 号：R450[医药卫生—治疗学]