Enhancement Effect of 2, 3-Dimethyl Maleic Acid on Luminol Chemiluminescence Reactions and Its Application in Detection of Sequence-Specific DNA Related to Hepatitis B Virus.

2, 3-dimethyl maleic acid (DMMA) was found to enhance luminol-H2O2 chemiluminescent (CL) reactions, among which the strongest enhancement effect was observed by using polyethyleneimine-templated gold nanoclusters (PEI-Au NCs) as the catalyst. With the addition of DMMA, the CL signal of the PEI-Au NCs-catalyzed luminol-H2O2 reaction enhanced about 630-fold, and a flash-type CL profile was obtained. Mechanism studies showed that the luminophore was still 3-aminophthalate anions in the excited state (3-APA*), and superoxide radical (O2·-) played an important role during the CL process. Under the optimized experimental conditions, the lowest concentration of PEI-Au NCs can be detected was 0.168 nM which was 82-fold lower than that without an enhancer. Furthermore, the catalytic activity of biotinylated PEI-Au NCs in the DMMA-enhanced luminol system was similar to PEI-Au NCs, providing a good opportunity for the development of CL bioanalysis platforms using PEI-Au NCs as the label. Thus, the DMMA-enhanced luminol-H2O2 system was applied to the CL detection of sequence-specific DNA related to the hepatitis B virus (HBV) using PEI-Au NCs as the label. The CL platform exhibited linearly enhanced CL response with the increasing amount of target DNA ranging from 0.0025 to 0.5 pmol. As low as 0.002 pmol of HBV DNA could be sensitively detected, which was superior to the previously reported methods.