DETECTION OF CAMPYLOBACTER PCR DNA BY HYBRIDIZATION WITH A MICROTITERPLATE IMMOBILIZED RNA PROBE

A microtiter plate hybridization system for the detection of Campyloba cter polymerase chain reaction (PCR) amplified DNA was developed using immobilized RNA probes. Digoxigenin-labelled or unlabelled PCR DNA wa s hybridized with immobilized RNA probe followed by immunoenzymatic as say of the hybrids using either anti-digoxigenin antibody or a monoclo nal anti-RNA:DNA hybrid antibody detection system. The application of probes specific for thermophilic Campylobacter and for Campylobacter j ejuni is demonstrated. With 10 ng of RNA immobilized in the microtiter plate, it was possible to accomplish hybridization in a microtiter we ll with a sensitivity level of 10 pg for the detection with digoxigeni n-labelled DNA and 25 pg with the monoclonal antibody RNA:DNA hybrid. This rapid and simple hybridization and immunoenzymatic detection syst em should be broadly applicable and will facilitate the assay of PCR-a mplified DNA from a variety of food and clinical samples.