Production of polyclonal and monoclonal antibodies for specific detection of nitrosation-proficient denitrifying bacteria in biological fluids

Polyclonal and monoclonal antibodies were raised against the nitrosating enzyme previously isolated and purified from a denitrifying bacteria Pseudomonas aeruginosa. Using the polyclonal antibodies, a preliminary ELISA test was set up which allowed the detection of the nitrosating enzyme in 2 ml urine samples with a minimal total bacteria count of greater than or equal to 10(5) cells/ml. The use of such a rapid immunological screening test in clinical settings should ascertain whether individual subjects at higher risk for cancer of the stomach or bladder harbour more nitrosation-proficient microorganisms in their microflora. The availability of monoclonal antibodies provides a tool for studying the mechanisms of bacteria-mediated nitrosamine formation from precursor amines.