Development of novel methods to identify biomolecules based on the conformational change of hairpin probe

Abstract

Since the increase of average human lifespan and population aging, the importance of early diagnostics has been raised and the market of in vitro diagnostic (IVD) has been increased consistently in recent 10 years. We herein described novel detection methods for biomolecule by utilizing the conformational change of hairpin probe induced by target molecule. In this study, we devised a detection system for target protein and activity of ribonuclease H (RNase H) based on the DNA polymerase activity, which is regulated by the combination of hairpin structured DNA aptamer and target recognition strand. Furthermore, by introducing the self-priming re-action of hairpin probe to isothermal nucleic acid amplification, we successfully simplified the reaction mecha-nism that is commonly complicated in isothermal amplification methods and developed highly sensitive detec-tion system for target nucleic acid. Since the devised systems are available for quantitative analysis of target molecules in a real-time manner, it is expected that the developed system would be a potential sensing tool in the field of IVD testing.