CHEMILUMINESCENT DEOXYOLIGONUCLEOTIDE PROBES FOR THE RAPID DETECTION OF INTACT COLIFORM BACTERIA AND TOTAL BACTERIA

BECAUSE BACTERIAL DISEASES ARE FREQUENTLY TRANSMITTED THROUGH CONTAMINATED WATER, THE DETECTION OF MICROBIOLOGICALCONTAMINATION IS IMPORTANT, TO ENSURE SANITARY WATER SUPPLIES. CURRENT TESTS ARE LABOR INTENSIVE AND TAKE SEVERAL DAYS TO CONFIRM CONTAMINATION LEVELS. THIS PROJECT WILL DEMONSTRATE THE FEASIBILITY OF A UNIQUE, RAPID, WATER-QUALITY TEST FOR BACTERIA. PHASE I FOCUSES ON THE DEVELOPMENT OF SPECIALIZED CHEMILUMINESCENT DEOXYOLIGONUCLEOTIDE PROBES. THESE DNA PROBES WILL COMPLEMENT UNIVERSAL BACTERIAL, COLIFORM-GROUP, AND E. COLI-SPECIFIC RIBOSOMAL RNA (RRNA) SEQUENCES AND, AS A RESULT, WILL HYBRIDIZE ONLY WHEN ONE OF THESE SEQUENCES IS PRESENT. THUS, IT WILL BE POSSIBLE RAPIDLY TO DETECT BACTERIAL CONTAMINATION. TO PRODUCE A VISIBLE RESPONSE, CHEMILUMINESCENT AGENTS WILL BE COVALENTLY LINKED TO THE DEOXYOLIGONUCLEOTIDE PROBES. THE PROBES WILL BE SHORT-15 TO 25 BASES LONG-ALLOWING INTACT CELLS TO BE PROBED WITHOUT FIRST EXTRACTING THE TARGET RRNA. TO MEASURE AND QUANTITATE THE CHEMILUMINESCENT EMISSION, A NUMBER OF COMMERCIALLY AVAILABLE LIGHT DETECTORS, INCLUDING FIELD-PORTABLE UNITS CURRENTLY IN DEVELOPMENT BY THE FIRM, WILL BE TESTED. USING THESE NEW PROBES, THE NUMBER OF BACTERIA IN WATER SAMPLES SHOULD BE DETERMINED IN AS LITTLE AS ONE HOUR. BECAUSE BACTERIAL DISEASES ARE FREQUENTLY TRANSMITTED THROUGH CONTAMINATED WATER, THE DETECTION OF MICROBIOLOGICALCONTAMINATION IS IMPORTANT, TO ENSURE SANITARY WATER SUPPLIES. CURRENT TESTS ARE LABOR INTENSIVE AND TAKE SEVERAL DAYS TO CONFIRM CONTAMINATION LEVELS. THIS PROJECT WILL DEMONSTRATE THE FEASIBILITY OF A UNIQUE, RAPID, WATER-QUALITY TEST FOR BACTERIA. PHASE I FOCUSES ON THE DEVELOPMENT OF SPECIALIZED CHEMILUMINESCENT DEOXYOLIGONUCLEOTIDE PROBES. THESE DNA PROBES WILL COMPLEMENT UNIVERSAL BACTERIAL, COLIFORM-GROUP, AND E. COLI-SPECIFIC RIBOSOMAL RNA (RRNA) SEQUENCES AND, AS A RESULT, WILL HYBRIDIZE ONLY WHEN ONE OF THESE SEQUENCES IS PRESENT. THUS, IT WILL BE POSSIBLE RAPIDLY TO DETECT BACTERIAL CONTAMINATION. TO PRODUCE A VISIBLE RESPONSE, CHEMILUMINESCENT AGENTS WILL BE COVALENTLY LINKED TO THE DEOXYOLIGONUCLEOTIDE PROBES. THE PROBES WILL BE SHORT-15 TO 25 BASES LONG-ALLOWING INTACT CELLS TO BE PROBED WITHOUT FIRST EXTRACTING THE TARGET RRNA. TO MEASURE AND QUANTITATE THE CHEMILUMINESCENT EMISSION, A NUMBER OF COMMERCIALLY AVAILABLE LIGHT DETECTORS, INCLUDING FIELD-PORTABLE UNITS CURRENTLY IN DEVELOPMENT BY THE FIRM, WILL BE TESTED. USING THESE NEW PROBES, THE NUMBER OF BACTERIA IN WATER SAMPLES SHOULD BE DETERMINED IN AS LITTLE AS ONE HOUR.