An amplification technology for improving sensitivity when measuring components in biological samples.

Abstract

A new technology for improving the sensitivity in measuring components in biological samples is described. The method is based on the use of spherical microbeads (detection beads) which contain a large amount of immobilized enzyme and a reagent with biospecific affinity for the component to be detected. These microbeads have been used in a 'sandwich reaction' for visualization of P-fimbriated Escherichia coli which has a known receptor structure (Gal(alpha 1-4)Gal(beta)). In the initial step the bacteria were enriched on a solid support (e.g., a plastic film or beads (greater than 150 microns)) to which the receptor structure had been covalently bound. In the next step detection beads coupled with enzyme and receptor structure were added and finally a chromogenic substrate for the enzyme was used for visualization. A sensitivity of 10(5) bacteria/ml was reached. The detection beads are of general utility and might be useful for the detection of lectins on other pathogens.