a tissue biopsy-based epigenetic multiplex pcr assay for prostate cancer detection组织biopsy-based后生多重pcr检测前列腺癌检测.pdfVIP

Van Neste et al. BMC Urology 2012, 12:16 /1471-2490/12/16 TECHNICAL ADVANCE Open Access A tissue biopsy-based epigenetic multiplex PCR assay for prostate cancer detection 1 1 2 1 1 3 Leander Van Neste , Joseph Bigley , Adam Toll , Gaëtan Otto , James Clark , Paul Delrée , Wim Van Criekinge1 and Jonathan I Epstein2* Abstract Background: PSA-directed prostate cancer screening leads to a high rate of false positive identifications and an unnecessary biopsy burden. Epigenetic biomarkers have proven useful, exhibiting frequent and abundant inactivation of tumor suppressor genes through such mechanisms. An epigenetic, multiplex PCR test for prostate cancer diagnosis could provide physicians with better tools to help their patients. Biomarkers like GSTP1, APC and RASSF1 have demonstrated involvement with prostate cancer, with the latter two genes playing prominent roles in the field effect. The epigenetic states of these genes can be used to assess the likelihood of cancer presence or absence. Results: An initial test cohort of 30 prostate cancer-positive samples and 12 cancer-negative samples was used as basis for the development and optimization of an epigenetic multiplex assay based on the GSTP1, APC and RASSF1 genes, using methylation specific PCR (MSP). The effect of prostate needle core biopsy sample volume and age of formalin-fixed paraffin-embedded (FFPE) samples was evaluated on an independent follow-up cohort of 51 cancer- positive patients. Multiplexing affects copy number calculations in a consistent way per assay. Methylation ratios are therefore altered compared to the respective singleplex assays, but the correlation with patient outcome remains equivalent. In addition, tissue-biopsy samples as small as 2