DNA PROBES FOR THE DIAGNOSIS OF BACTERIAL VAGINOSIS

THE ROLE OF FIVE BACTERIA AND A MYCOPLASMA AS THE ETIOLOGIC AGENTS OF BACTERIAL VAGINOSIS (BV) IS SUPPORTED BY GROWING SCIENTIFIC DATA. STANDARD METHODS ARE INADEQUATE FOR THE ROUTINE DIAGNOSIS OF THIS DISEASE. EIGHT HUNDRED TO 1,000 NUCLEOTIDES OF THE 16S RIBOSOMAL RNA (RRNA) OF EACH BV MICROORGANISM WILL BE SEQUENCED. SHORT DNA PROBES WILL BE SYNTHESIZED TO DETECT THE 16S RRNA OF EACH BV MICROORGANISM.32P-LABELED DNA PROBES WILL BE HYBRIDIZED AGAINST HIGH-MOLECULAR-WEIGHT RNA AND DNA ISOLATED FROM INDIVIDUAL PURE CULTURES. HYBRIDIZATION SPECIFICITY OF EACH DNA PROBE WILL BE EVALUATED IN TETRAETHYLAMMONIUM CHLORIDE AND TETRAMETHYLAMMONIUM CHLORIDE SOLUTIONS TO ABOLISH THE PREFERENTIAL MELTING OF A-T VERSUS G-C BASE PAIRS IN DNA HYBRIDS, YIELDING HYBRIDIZATIONS THAT ARE ONLY CHAIN-LENGTH DEPENDENT. IN THESE SOLUTIONS, MIXTURES OF PROBES OF THE SAME LENGTH, BUT DIFFERING IN SEQUENCE, WILL EXHIBIT IDENTICAL HYBRIDIZATION PROPERTIES. THESE SALT SOLUTIONS WILL ALSO ALLOW SIMULTANEOUS USE OF MULTIPLE PROBES (OF THE SAME LENGTH) AND WILL ALLOW THE DEVELOPMENT OF A UNIVERSAL HYBRIDIZATION FORMAT. THE LONG-TERM OBJECTIVE IS TO PROVIDE A RAPID AND ACCURATE DIAGNOSTIC TEST FOR BV.