No abstract

A novel colorimetric assay method based on enzyme-induced metallization has been proposed for detection of alkaline phosphatase (ALP), and it was further applied to highly sensitive detection of avian influenza virus particles coupled with immunomagnetic separation. The enzyme-induced metallization-based color change strategy combined the amplification of the enzymatic reaction with the unique optical properties of metal nanoparticles (NPs), which could lead to a great enhancement in optical signal. The detection limit for ALP detection was 0.6 amol/50 μL which was 4-6 orders of magnitude more sensitive than other metal NP-based colorimetric methods. Moreover, this technique was successfully employed to a colorimetric viral immunosensor, which could be applied to complex samples without complicated sample pretreatment and sophisticated instruments, and a detection limit as low as 17.5 pg mL(-1) was achieved. This work not only provides a simple and sensitive sensing approach for ALP and virus detection but also offers an effective signal enhancement strategy for development of a highly sensitive nonaggregation metal NP-based colorimetric assay method.

Single molecule electrochemistry (SME) has gained much progress in fundamental studies, but it is difficult to use in practice due to its less reliability. We have solved the reliability of single molecule electrochemical detection by integration of digital analysis with efficient signal amplification of enzyme-induced metallization (EIM) together with high-throughput parallelism of microelectrode array (MA), establishing a digital single molecule electrochemical detection method (dSMED). Our dSMED has been successfully used for alkaline phosphatase (ALP) detection in the complex sample of liver cancer cells. Compared to direct measurement of the oxidation current of enzyme products, EIM can enhance signals by about 100 times, achieving signal-to-background ratio high enough for single molecule detection. The integration of digital analysis with SME can further decrease the detection limit of ALP to 1 aM relative to original 50 aM, enabling dSMED to be sensitively, specifically and reliably applied in liver cancer cells. The presented dSMED is enormously promising in exploring physical and chemical properties of single molecules, single biomolecular detection, or single-cell analysis.