The characterization of Gardnerella vaginalis DNA using non-radioactive DNA probes.
DNA restriction profiles of various Gardnerella vaginalis isolates, generated by BamHI, EcoRI, PstI and other restriction enzymes, varied considerably. Only a few DNA fragments were identified as common in ethidium bromide fluorescence profile and Southern-blot hybridization patterns (employing a digoxigenin-labelled G. vaginalis DNA probe and an enzyme-linked immunoassay detection method). While the efficiencies of Southern-blot hybridization appeared inconsistent, in dot-blot assays, DNA from each isolate hybridized readily, enabling the detection of at least 10 ng DNA. A 5.7-kb DNA fragment from G. vaginalis ATCC 14018 genomic library, cloned in the BamHI site of pBR322, could replace the total genomic DNA probe. This specific DNA fragment was present in different sizes in 12 analysed G. vaginalis strains, describing a restriction fragment length polymorphism. In control studies, none of the DNA from bacteria other than G. vaginalis (including some genitourinary tract residents) hybridized with the G. vaginalis total or specific DNA probes. Non-radioactive G. vaginalis DNA probes can thus form the basis of a useful detection method for further studies of this organism.
Nath
K
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Department of Biology, Long Island University, Brookville, NY 11548.
Choi
D J
DJ
Devlin
D
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D015139:Blotting, Southern; D015342:DNA Probes / Q000032:analysis*; D004269:DNA, Bacterial / Q000032:analysis*; D005260:Female; D006194:Gardnerella vaginalis / Q000235:genetics*; D006801:Humans; D007124:Immunoenzyme Techniques; D066298:In Vitro Techniques; D015183:Restriction Mapping
D015342:DNA Probes; D004269:DNA, Bacterial