Novel polystyrene microspheres-based chemiluminescence technology for the amplified detection of thrombin

A novel chemiluminescence (CL) immunoassay for the amplified determination of thrombin is proposed, by using two aptamer sequences as recognition elements, 96-well microplate as a reaction carrier, polystyrene microspheres as an amplification platform and horseradish peroxidase (HRP) as a CL label. The results show that the amplified CL technique couples the high sensitivity of CL analysis with effective discrimination against unwanted proteins such as IgG, IgM, IgA, IgE and IFN. An excellent linearity from 7.8 to 250 pM with the lowest detection limit of 3.9 pM was reached for the amplified technique whereas a good linear correlation from 0.98 to 30 nM with a detection limit of 0.46 nM was achieved for the nonamplified method, which was more than 100-fold less sensitive than using the amplified approach. Overall, this proposed detection scheme is time-effective with high sensitivity and throughout. These features, as well as its operation convenience, make it a promising alternative to thrombin detection methods.