The proposed work will result in a simple-to-use and inexpensive prototype test kit for the rapid and accurate differentiation of the major pathogens responsible for vaginitis. Vaginitis is among the most common conditions for which women seek medical attention. This kit will be used in the doctor's office, clinical and laboratory settings. For this test, a panel of synthetic oligonucleotides will be developed that specifically detects the abundant 16S and 23S ribosomal RNAs released from target microorganisms. This will be accomplished using a sandwich assay format in which the organism-specific oligonucleotides will be covalently coupled to a solid support and allowed to hybridized with lysis-hybridization reagent. The resulting captured rRNAs will be detected by the addition of a second set of ligonucleotides that will hybridize with the conserved sequences present in the rRNA. This second set of DNA probes will be coupled with a highly sensitive clorimetric signal production system. Finally, the prototype kit thus developed will be evaluated using clinical specimens.
| Sheiness, D; Dix, K; Watanabe, S et al. (1992) High levels of Gardnerella vaginalis detected with an oligonucleotide probe combined with elevated pH as a diagnostic indicator of bacterial vaginosis. J Clin Microbiol 30:642-8 |
