Research Project
3436662
For the past quarter of a century, one of the assumed etiological agents of bacterial vaginosis or nonspecific vaginitis has been masquerading under the guise of a Haemophilus or a Corynebacterium. However, within the last two years this bacterium has emerged to be a single novel species, now known as Gardnerella vaginalis. The role of G. vaginalis in nonspecific vaginitis is controversial due to several difficulties, lack of a convenient diagnosis method being one of them. Very little is known about this bacteria besides its etiology and probable taxonomy. The primary aim of this proposal is to construct a superior, i.e. rapid, simple and reliable detection method for G. vaginalis utilizing methods of recombinant DNA technology. G. vaginalis is variably Gram - staining small bacillus. DNA from such organism will be isolated after lysozyme treatment of cell wall and detergent lysis of the cell, removal of protein by denaturation, and final purification from a cesium chloride equilibrium genomic library will be constructed in Escherichia coli utilizing plasmid pBR325 as the vector. Amplified Gardnerella DNA fragments will be utilized in situ hybridization studies on cytological smears with the presence of "Clue cells" or specimens that are known to be infected by Gardnerella from cytological studies. The proposed approach is new and novel to G. vaginalis, an organism associated not only with bacterial vaginosis but with postpartum endometritis, amniotic fluid infection, neonatal sepsis and various genitourinary infections in men. We wish to carry on future studies on the case of pathogenicity, since 40 to 60% women and 10 to 15% men show no adverse effect. Are there biotypes of Gardnerella that may be reflected on the genomic level? There are curious observations of early stages of cytological transformation of cervico-vaginal squamous epithelial in the presence of large number of Gardnerella. Our future studies will involve the possible role of G. vaginalis in squamous atypia leading to a possible squamous dysplasia.
