【摘要】 目的建立Bcl-2/IgH融合基因实时定量PCR检测方法。方法以淋巴瘤患者Bcl-2/IgH融合基因的纯化DNA作为标准品,淋巴瘤患者骨髓和外周血为样本,建立SYBR GreenⅠ荧光染料实时定量PCR(Real-time Quantitative PCR,RQ-PCR),并对方法的灵敏性、稳定性、重复性进行测定。结果构建的RQ-PCR方法检测Bcl-2/IgH融合基因的敏感性达10-7水平;标准曲线的斜率和相关系数分别为-3.13,0.99;管间变异和批间变异分别为6.54%,6.73%,1.90%和3.59%,6.30%,5.49%。结论建立的SYBR GreenⅠ荧光染料RQ-PCR方法灵敏,标准曲线的相关性好,方法的稳定性和重复性较好。更多还原

【Abstract】 Objective To establish a real-time quantitative polymerase chain reaction (RQ-PCR) method for detection of Bcl-2/IgH fusion gene in lymphoma. Methods SYBR Green RQ-PCR was constructed by using the purified DNA from Bcl-2/IgH-positive lymph node cells as standard sample and those from bone marrow(BM)and/or peripheral blood(PB)of lymphoma patients as samples. The sensitivity,stability and repeatability of this method were determined.Result The Result showed the sensitivity of the established real-time quantitative PCR for detecting Bcl-2/IgH fusion gene was 10-7 level. The slope and coefficient correlation were-3.13 and 0.99 respectively. The coefficient variation(CV) among tubes and batches were 6.54%,6.73%,1.90% and 3.59%,6.30%,5.49% respectively. Conclusions The standard curve had well linear relationship and the established SYBR Green real-time quantitative PCR method has the advantage of high sensitivity,good stability and reproducibility.更多还原