Non-enzymatic aqueous peroxyoxalate chemiluminescence immune detection using a CCD camera and a CMOS device

A new method for non-enzymatic aqueous peroxyoxalate chemiluminescence (POCL) biomolecular detection using imaging chip-based devices has been developed. A water-soluble amide of oxalic acid was synthesized and used in the investigation and characterization of POCL immunodetection in an aqueous environment. Six fluorescent dyes commonly used in biological detection were tested, and the intensity of light generated from the aqueous POCL reactions was characterized in the liquid phase. Direct detection sensitivity comparisons between a standard fluorescent method and this POCL method were performed in both liquid and solid phases. Results showed that detection sensitivity using the POCL method is comparable to that of the fluorescent method. POCL biomolecular detection on a nitrocellulose membrane was also investigated using a charge-coupled device (CCD) camera. Again, POCL detection sensitivity proved to be comparable to that using the fluorescent detection method. In an application of aqueous POCL biomolecular detection, Staphylococcus aureus enterotoxin B (SEB) and its antibody were used to demonstrate immuno- and affinity detection. For further applications, such as DNA and protein arrays, simultaneous detection of biomolecules labelled with different fluorescent labels was investigated, using a complementary metal oxide semiconductor (CMOS) colour imaging chip.