摘要
目的 建立TaqMan实时荧光定量逆转录聚合酶链反应 (RT PCR)法检测新型趋化因子巨噬细胞炎症蛋白 2γ(MIP 2γ)mRNA表达水平 ,并对方法进行评估。方法 TaqMan实时荧光定量RT PCR检测MIP 2γmRNA的表达。结果 线性检测范围达 6个数量级 ,最低检测下限为 6× 10 2 拷贝 ,最高检测上限为 6× 10 7拷贝 ,批间及批内变异 <12 2 %。正常Balb/c小鼠心脏、肝脏、肾脏组织均可检测到MIP 2γmRNA表达 ,以心脏表达水平最高。结论 采用TaqMan实时荧光定量RT PCR检测MIP 2γmRNA表达水平准确、特异、灵敏、快速、操作简便 ,具有临床推广价值。
Objectives To develop and evaluate a real-time fluorescent quantitative RT-PCR method for the detection of a novel CXC chemokine macrophage inflammatory protein-2γ(MIP-2γ)mRNA expression based on TaqMan technique.Method The expression of MIP-2γ mRNA was detected by TaqMan real time fluorscet quantitative RT-PCR. Results The dynamic range of the assay varied from 102 to 107 copies. The intra- and inter-assay coefficient variation (CV) were less than <122%. MIP-2γ mRNA is expressed in heart, liver and kidney of normal Balb/c mice, with the greatest expression in heart.Conclusion The detection of MIP-2γ mRNA expression by real time fluorescent quantitative RT-PCR based on TaqMan technique is more accurate,specific, sensitive and time-saved, which is suitable for use in clinical laboratory.
出处
《中华检验医学杂志》
CAS
CSCD
北大核心
2003年第5期290-292,共3页
Chinese Journal of Laboratory Medicine
