淋巴瘤Bcl-2/IgH融合基因实时定量PCR检测方法的研究

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。

Establishment of a Real-time Quantitative Polymerase Chain Reaction for Detection of Bcl-2/IgH Fusion Gene in lymphoma

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.