ATP Detection Research Based On Nucleic Acid Aptamer And DNA Chip

Background: Nucleic acid aptamer is a artificial single-strandedoligonucleotides,which can combined with the target with high specificityand affinity. It was screened out from the big capacity of artificial buildingoligonucleotide library by systematic evolution of ligands by exponentialenrichment(SELEX). Its function is similar to the antibody, compared withantibody, nucleic acid aptamer has many advantages.Using the SELEXtechnology,different nucleic acid aptamers have been selected and theseaptamers have been widely used in drug development, food sanitation,basic medical research, disease diagnosis, clinical treatment and so on. Atpresent, the research of nucleic acid aptamer in material detection areincreasing, and the detecting techniques based on nucleic acid aptamer areconstantly emerging. Nucleic acid aptamer is playing an important role inanalysis modes of flow cytometry, biosensors, microfluidic cell separation,fluorescence polarization, molecular beacon, electrochemical impedancespectroscopy and histological examination. Objective: We built a high sensitivity and specificity detectionmethod for ATP based on nucleic acid aptamer,DNA chips and nanogoldlabelling technology.Methods:1. ATP nucleic acid aptamer was taken as recognitionmolecule for ATP detection. The sequence of ATP nucleic acid aptamerwas obtained through the literature, designed the complementary nucleicacid sequence of ATP nucleic acid aptamer and fixed it to the aldehydegroup as capture probe.2. Established and optimized the ATP detectionmethod based on nucleic acid aptamer and DNA chip. Including: thecapture probe fixed concentration, the hybridization time of capture probeand biotin-modified ATP nucleic acid aptamer, the response time of ATPand its nucleic acid aptamer, the time of silver enhancement, the pH valueand Mg2＋concentration of ATP and its nucleic acid aptamer reaction.3.Under the optimized conditions, we evaluated the sensitivity of this methodby testing the different concentrations of ATP, respectively, and evaluatedthe specificity of this method by testing ATP, ATP similar materials,and ATP different materials, respectively,.Results: The fixed efficiency reached the highest when theconcentration of the capture probe for0.6μM. The detection performanceof the method achieved the best under the condition of55min forhybridization time,45min for ATP reaction time,7.80of pH and6mM Mg2＋for ATP and its nucleic acid reaction,5.5min for silver enhancement time. Tested the different concentrations of ATP under the optimal conditions,we found that the relative intensity of the greyscale was proportional to thelogarithmic value of ATP concentration over a5-decade range from10-14Mto10-9M with a linear correlation coefficient of0.996. We tested the TTP,UTP, CTP, GTP, BSA and ATP respectively, the results demonstrated thatthe proposed method has high specificity for ATP detection.Conclusion: This study built a ATP detection method based onnucleic acid aptamer, DNA chips and nanogold labelling technology withhigh specificity and sensitivity, and could provide a fresh perspective forthe detection of other materials.