Rapid and specific detection of bacteria using bioluminescence

Abstract

This paper describes rapid methods for the detection of very low numbers of bacteria. Specificity was obtained by the use of antibodies in immunomagnetic separation, a bacteriophage to allow targeted cell lysis, or a combination of both. The amplified endpoint assay used cell-derived adenylate kinase to convert added adenosine diphosphate (ADP) to adenosine triphosphate (ATP) which could then act as a substrate for the firefly bioluminescence reaction. Specific and non-specific assays were evaluated using Escherichia coli O157 as the test organism. Limits of detection of around 10 2 cells ml −1 could be obtained with an upper end cut-off of around 10 7 cells ml −1. Depending on the level of specificity offered, the assays took from 5 min to just under 1 h.