摘要
目的前列腺癌抗原3(prostate cancer antigen 3,PCA3)基因表达具有前列腺癌特异性,文中建立荧光实时定量PCR(FQ-RT-PCR)检测PCA3评分的方法。方法分别以LNCap和PC-3细胞株为阳性和阴性对照,用TaqMan探针建立检测PCA3和前列腺特异性抗原(prostate specific antigen,PSA)mRNA的荧光实时定量PCR方法,评价特异性、重复性和计算扩增效率,建立PCA3评分计算公式。结果检测PCA3和PSA mRNA的批内变异系数分别为1.19%、1.09%、0.84%和0.06%、0.71%、0.79%,批间变异系数分别为1.03%和0.56%;扩增效率分别为0.98和1.00;以2-ΔCt表示PCA3评分。结论荧光实时定量PCR检测PCA3评分方法的特异性好,重复性高。
Objective Prostate cancer gene 3(PCA 3) is specific for prostate cancer.This paper is to establish a real-time fluorescence quantitative polymerase chain reaction(FQ-RT-PCR) method for determining the PCA3 score.Methods LNCap and PC-3 cell lines were used as positive and negative controls,respectively,and a real-time fluorescence quantitative PCR method was established with the TaqMan probe for the detection of PCA3 and PSA mRNA,evaluate its specificity,reproducibility and amplification efficiency,and then develop a formula for determining the PCA3 score.Results The intra-assay variation coefficients of PCA3 were 1.19%,1.09% and 0.84%,and those of PSA mRNA were 0.06%,0.71% and 0.79%;the inter-assay variation coefficients of PCA3 and PSA mRNA were 1.03% and 0.56%,respectively.And the amplification efficiencies of PCA3 and PSA mRNA were 0.98 and 1.00,respectively.The PCA3 score was expressed by 2-ΔCt.Conclusion FQ-RT-PCR has high specificity and reproducibility for the determination of the PCA3 score.
出处
《医学研究生学报》
CAS
2011年第6期569-572,共4页
Journal of Medical Postgraduates
基金
南京军区南京总医院青年基金(Q2008059)
关键词
前列腺癌抗原3
前列腺特异性抗原
前列腺癌
荧光实时定量PCR
Prostate cancer antigen 3
Prostate specific antigen
Prostate cancer
Real-time fluorescence quantitative polymerase chain reaction
