Detection Methods And Clinical Application Of CEA-mRNA By Real-Time Flurogenic Quantitative Reverse Transcriptase Polymerase Chain Reaction In Colorectal Cancer Patients

Objective To develop a real-time quantitative RT-PCR method for detection of CEA mRNA ,and with the method to detect the CEA mRNA expression of cancer tissues , lymph nodes and peripheral blood in Colorectal Cancer patients. Exploring the functions and clinical significance in patients of Colorectal Cancer.Methods With the established method, we detected the CEA mRNA expression of tumor tissues, lymph nodes and peripheral blood in 56 colorectal cancer patients,30 colon benign patients and 30 normal control,we also detected CEA protein expression of the three groups people and analyzed the relation of CEA mRNA with different Dukes stages , pathology differentitation,and compared the CEA mRNA expression changes before surgery and after surgery, and analyzed the correlation between CEA mRNA and CEA protein.Results1. Using SYBR Green I as report dye and GAPDH as inner reference, a real-time quantitative RT-PCR method was established. The amplification efficiencies for CEA and GAPDH of the real-time quantitative RT-PCR method were 0.99 and 0.98 respectively, inter-coefficient of variation were 2.0% and 0.8% respectively, intra--coefficient of variation were 3.2% and 3.9% respectively. The electrophoresis and melting curve analysis of gene amplification product showed that the method for CEA mRNA quantification with SYBR...