Isolation of a species-specific DNA probe for Neisseria gonorrhoeae using a novel technique particularly suitable for use with closely related species displaying high levels of DNA homology

Molecular and Cellular Probes
J J DoneganH L Yang

Abstract

The use of nucleic acid probes has become an increasingly common method for detecting pathogenic micro-organisms in clinical specimens. In the course of our efforts to isolate species-specific DNA probes for bacterial pathogens, we encountered a special problem with regard to Neisseria gonorrhoeae. As a consequence of the high degree of DNA homology that this organism displays with its nearest relative, Neisseria meningitidis, the isolation of such probes could not be readily achieved. We therefore developed a novel method of probe isolation which overcomes this problem. This methodology relies upon the application of a 'sandwich' hybridization assay to screen an M13 'shotgun' library derived from N. gonorrhoeae genomic DNA. For this, genomic DNA from N. gonorrhoeae and N. meningitidis was immobilized on nitrocellulose filters and probed with recombinant phage DNA from candidate clones. Those clones which had hybridized to target sequences were then detected using labelled vector sequences in a second hybridization step. By this means we obtained a numerical assessment of the degree of specificity of candidate clones for the target organism as compared to one or more related species. Using this technique we have isolated three ...Continue Reading

References

Oct 1, 1975·Proceedings of the National Academy of Sciences of the United States of America·M Grunstein, D S Hogness
Mar 1, 1987·Molecular and Cellular Probes·R P Viscidi, R G Yolken
Sep 1, 1987·Molecular and Cellular Probes·R N PickenH L Yang
Jan 24, 1981·Nucleic Acids Research·J MessingP H Seeburg

❮ Previous
Next ❯

Citations

Jul 1, 1992·European Journal of Clinical Microbiology & Infectious Diseases : Official Publication of the European Society of Clinical Microbiology·R J CanoR E Klem

❮ Previous
Next ❯

Related Concepts

Related Feeds

Bacteriophage: Phage Therapy

Phage therapy uses bacterial viruses (bacteriophages) to treat bacterial infections and is widely being recognized as an alternative to antibiotics. Here is the latest research.