Netlike hybridization chain reaction assembly of DNA nanostructures enables exceptional signal amplification for sensing trace cytokines

The monitoring and detection of molecular biomarkers play crucial roles in disease diagnosis and treatment. In this work, we proposed a target-responsive netlike hybridization chain reaction (nHCR) DNA nanostructure construction method, which can offer an exceptional signal enhancement, for highly sensitive fluorescence detection of cytokine, interferon-gamma (IFN-gamma). The presence of the target cytokine can lead to the conformational change of the aptamer recognition hairpin probes and the liberation of the nHCR initiator strands, which further trigger the nHCR process between two dye-labeled and double hairpin-structured probes to form netlike DNA nanostructures. The formation of the DNA nanostructures brings the dyes into close proximity, resulting in significantly amplified fluorescence resonance energy transfer signals for sensitive and enzyme-free detection of IFN-gamma. The present method has a detection limit of 1.2 pM and a dynamic linear range of 5 to 1000 pM for IFN-gamma detection. Besides, with the high specificity of the aptamer probe and the significant signal amplification of the nHCR, such an IFN-gamma detection strategy shows excellent selectivity and high sensitivity, which can be potentially applied to detect IFN-gamma in human serums. With such a demonstration of the detection of IFN-gamma, this proposed method can be extended for detecting different types of biomolecules.