摘要
目的:建立荧光实时定量PCR技术,检测非小细胞肺癌组织与外周血RRM1和ERCC1及BRCA1基因表达水平。方法:分别构建RRM1、ERCC1和BRCA1及管家基因β-actin质粒标准品,以SYBR荧光实时定量PCR分析,制备标准曲线,对非小细胞肺癌组织与外周血中RRM1、ERCC1和BRCA1及管家基因β-actin的mRNA进行检测。结果:标准曲线呈良好的线性关系。标准品的熔解曲线均呈单峰,特异性良好,说明基本无非特异性扩增。结论:所建SYBR荧光实时定量PCR方法操作简便,费用低,特异性好,准确度、灵敏度高,为后续研究构建了理想的平台。
Objective: To develop a method for the detection of RRM1,ERCC1 and BRCA1 gene expression by SYBR real-time fluorescent quantitative PCR in non-small cell lung cancer tissues and peripheral blood.Methods: The plasmid standard of RRM1,ERCC1,BRCA1 and β-actin genes was constructed.SYBR real-time PCR was performed,and the standard curve was established.The expressions of RRM1,ERCC1 and BRCA1 mRNA in non-small cell lung cancer tissues and peripheral blood were detected.Results: The standard curve presented linearity.The liquate curves of standard gene were all single apex,indicating that a good specificity was obtained.Conclusion: The developed SYBR real-time fluorescent quantitative PCR has advantage of convenient operation,low cost,good specificity and high veracity.
出处
《浙江大学学报（医学版）》
CAS
CSCD
北大核心
2010年第6期628-633,共6页
Journal of Zhejiang University(Medical Sciences)
基金
浙江省科技计划资助项目(2009C33165)
浙江省医药卫生科学研究基金计划项目(2008A060)
