CAJ | 학술논문

目的建立一套快速、灵敏和特异的人抗黏液蛋白A(MxA)基因启动子中干扰素刺激应答元件-88/-123位多态性检测体系,为合理应用IFN-α治疗慢性乙型肝炎提供分子生物学检测手段.方法对经IFN-α治疗的患者进行HBV DNA、HBV基因分型及MxA基因启动子中干扰素刺激应答元件-88/-123位多态性位点检测,确定基因多态性与IFN-α治疗效果之间的关系.通过各种条件优化实验,建立MxA基因启动子中干扰素刺激应答元件-88/-123位多态性荧光PCR检测体系,并通过与DNA序列分析法检测结果的对比,验证荧光PCR检测体系的灵敏性及特异性,从而对该体系的临床适用性进行初步评估.采用卡方检验计算P值、回归分析法计算对比率和95%可信区间.结果 MxA基因启动子干扰素刺激应答元件中-88位为G/T杂合型和-123位为C/A杂合型者皆为IFN-α敏感型,-88位为G/G纯合型和-123位为C/C纯合型者为IFN-α不敏感型.与DNA序列分析的金标准相比,荧光PCR检测的符合率为99.65%.结论荧光PCR检测体系,可灵敏、快捷地检测患者MxA基因多态性位点.
목적 건립일투쾌속、령민화특이적인항점액단백A(MxA)기인계동자중간우소자격응답원건-88/-123위다태성검측체계,위합리응용IFN-α치료만성을형간염제공분자생물학검측수단.방법 대경IFN-α치료적환자진행HBV DNA、HBV기인분형급MxA기인계동자중간우소자격응답원건-88/-123위다태성위점검측,학정기인다태성여IFN-α치료효과지간적관계.통과각충조건우화실험,건립MxA기인계동자중간우소자격응답원건-88/-123위다태성형광PCR검측체계,병통과여DNA서렬분석법검측결과적대비,험증형광PCR검측체계적령민성급특이성,종이대해체계적림상괄용성진행초보평고.채용잡방검험계산P치、회귀분석법계산대비솔화95%가신구간.결과 MxA기인계동자간우소자격응답원건중-88위위G/T잡합형화-123위위C/A잡합형자개위IFN-α민감형,-88위위G/G순합형화-123위위C/C순합형자위IFN-α불민감형.여DNA서렬분석적금표준상비,형광PCR검측적부합솔위99.65%.결론 형광PCR검측체계,가령민、쾌첩지검측환자MxA기인다태성위점.
Objective To establish a fluorescent polymerase chain reaction (PCR) method for rapid, sensitive and specific determination of -88/-123 polymorphisms in Myxovirus resistance protein A (MxA) gene promoter so as to provide molecular biology tool for optimized interferon-a treatment in chronic hepatitis B patients. Methods Hepatitis B virus (HBV) genotyping,serum HBV DNA level,and- 88/- 123 polymorphisms in MxA gene promoter of patients who had been treated with interferon-α were detected. The statistical analysis was done by using SPSS software to understand the relationship between MxA gene polymorphisms and interferon-α treatment. Afterwards, an optimal fluorescent PCR system was established to determine -88/-123 polymorphisms in MxA gene promoter. The sensitivity and the specificity of this system were confirmed by DNA sequencing. P-value of chi square test, odds ratios of regression analysis and 95% confidence intervals were employed. Results Patients with- 88 G/T and - 123 C/A in the interferon-stimulated response element in MxA gene promoter were interferon-α sensitive, while patients with - 88 GIG and - 123 C/C were not interferon-α sensitive. The coincidence rate of this system was 99.65% in comparison with DNA sequencing.Conclusion MxA gene polymorphisms could be rapidly and sensitively determined by this fluorescent PCR system.