Detect Intracellular Activity Of Tumor Markers Based On Isothermal Strand Displacement Reaction

Cancer, one of the most threatening diseases of the human being, incidence rate is becoming increasingly high as survival environment worsening. However, cure rate of cancer is very low. So it is a medical problem for early detection and treatment of the cancer. The earlier presence of cancer been found, the higher cure rate patient will have.Analysis and detection of active substance in tumour cells has a great significance for the prevention and treatment of cancer. This thesis reported new methods based on nanotechnology, DNA biosensor and fluorescenceassay technology for detection and analysis of active substance in tumour cells.The main research contents include the following aspects:1、Based on the extension reaction of a telomerase substrate(TS) primer in the presence of the telomerase, and the recessed 3’-hydroxyl termini of duplex DNA in the presence of Exonuclease-III(Exo-III).Using this scheme can detect the lowest concentration of commerical telomerase is 5.0 × 10-14 IU/mL, can detect 6 Hela and 10 Romas cells.2、Based on the polymerase chain reaction to open the intrinsic fluorescence hairpin probe signal amplification technique detection of telomerase activity. The lowest concentration of commerical telomerase is 5.0 × 10-10 IU/L and the telomerase activity in cell extracts equivalent to 4 HeLa cells was detected by this method.3、Based on the principle of equivalent temperature chain substitution reaction, a new type of DNA sensor was designed to detect ATP and glutathione simultaneously.Using this scheme can detect the lowest concentration of ATP is 2.0 × 10-8 mol/L andthe lowest concentration of GSH is 1.0 × 10-9 mol/L.