摘要
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HBVDNA、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。
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-α treatment in chronic hepatitis B patients. Methods Hepatitis B virus （HBV） genotyping,serum HBV DNA level, and- 88/- 123 polyrnorphisms 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 G/G 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.
出处
《中华传染病杂志》
CAS
CSCD
北大核心
2008年第10期580-584,共5页
Chinese Journal of Infectious Diseases
基金
广东省医学科学技术研究基金资助项目（A2005643）
深圳市科技计划重点项目（JH200505270415B）
深圳科技计划项目（2003～131）
