淋巴瘤Bcl-2/IgH融合基因实时定量PCR检测方法的研究  

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

作　　者：张学美[1] 徐明[2] 刘华[3] 王玉明[3] 李惠民[1] 李云涛[1] 

机构地区：[1]昆明医学院第一附属医院血液科 [2]昆明市第一人民医院血液肿瘤科,昆明650032 [3]昆明医学院第一附属医院检验科

出　　处：《中国医疗前沿》2009年第23期7-8,12,共3页China Healthcare Innovation

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。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.

关 键 词：BCL-2/IGH 融合基因 淋巴瘤 实时定量PCR 

分 类 号：R733.4[医药卫生—肿瘤]