Abstract
Phenotypic characterization of 19 Neisseria gonorrhoeae serovar IB-2 and 8 serovar IB-6 isolates by the combined use of auxotypes, serological characterization, and penicillin susceptibility testing indicated intraserovar genetic diversity. In the present study, we applied whole-cell repetitive element sequence-based PCR (rep-PCR) analysis which allows a rapid assessment of the clonal relationships of IB-2 and IB-6 isolates. DNA templates were prepared by boiling cells harvested directly from plate cultures, eliminating the need for time-consuming phenol extraction. Six different rep-PCR profiles were established among the 19 IB-2 isolates. Rep-PCR typing results had a good correlation with pulsed-field gel electrophoresis patterns. It is slightly less discriminatory than BglII-generated macrorestriction pattern analysis by pulsed-field gel electrophoresis. It is capable of discriminating epidemiologically related from epidemiologically unrelated IB-2 isolates. It should serve as a rapid and useful subtyping tool for epidemiologic investigations in which there is a predominance of major serovar groups.
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- Dasi M. A., Camarena J. J., Ledesma E., García R., Moreno F., Nogueira J. M., Hernández J. Random amplification of polymorphic DNA of penicillinase-producing Neisseria gonorrhoeae strains. Genitourin Med. 1993 Oct;69(5):404–405. doi: 10.1136/sti.69.5.404. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Geers T. A., Donabedian A. M. Comparison of broth microdilution and agar dilution for susceptibility testing of Neisseria gonorrhoeae. Antimicrob Agents Chemother. 1989 Feb;33(2):233–234. doi: 10.1128/aac.33.2.233. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Georghiou P. R., Doggett A. M., Kielhofner M. A., Stout J. E., Watson D. A., Lupski J. R., Hamill R. J. Molecular fingerprinting of Legionella species by repetitive element PCR. J Clin Microbiol. 1994 Dec;32(12):2989–2994. doi: 10.1128/jcm.32.12.2989-2994.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gill M. J. Serotyping Neisseria gonorrhoeae: a report of the Fourth International Workshop. Genitourin Med. 1991 Feb;67(1):53–57. doi: 10.1136/sti.67.1.53. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Poh C. L., Loh G. K., Tapsall J. W. Resolution of clonal subgroups among Neisseria gonorrhoeae IB-2 and IB-6 serovars by pulsed-field gel electrophoresis. Genitourin Med. 1995 Jun;71(3):145–149. doi: 10.1136/sti.71.3.145. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sandström E. G., Rudén A. K. Markers of Neisseria gonorrhoeae for epidemiological studies. Scand J Infect Dis Suppl. 1990;69:149–156. [PubMed] [Google Scholar]
- Sarafian S. K., Knapp J. S. Molecular epidemiology of gonorrhea. Clin Microbiol Rev. 1989 Apr;2 (Suppl):S49–S55. doi: 10.1128/cmr.2.suppl.s49. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Versalovic J., Kapur V., Mason E. O., Jr, Shah U., Koeuth T., Lupski J. R., Musser J. M. Penicillin-resistant Streptococcus pneumoniae strains recovered in Houston: identification and molecular characterization of multiple clones. J Infect Dis. 1993 Apr;167(4):850–856. doi: 10.1093/infdis/167.4.850. [DOI] [PubMed] [Google Scholar]
- Versalovic J., Koeuth T., Lupski J. R. Distribution of repetitive DNA sequences in eubacteria and application to fingerprinting of bacterial genomes. Nucleic Acids Res. 1991 Dec 25;19(24):6823–6831. doi: 10.1093/nar/19.24.6823. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Welsh J., McClelland M. Fingerprinting genomes using PCR with arbitrary primers. Nucleic Acids Res. 1990 Dec 25;18(24):7213–7218. doi: 10.1093/nar/18.24.7213. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Woods C. R., Jr, Versalovic J., Koeuth T., Lupski J. R. Analysis of relationships among isolates of Citrobacter diversus by using DNA fingerprints generated by repetitive sequence-based primers in the polymerase chain reaction. J Clin Microbiol. 1992 Nov;30(11):2921–2929. doi: 10.1128/jcm.30.11.2921-2929.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Woods C. R., Versalovic J., Koeuth T., Lupski J. R. Whole-cell repetitive element sequence-based polymerase chain reaction allows rapid assessment of clonal relationships of bacterial isolates. J Clin Microbiol. 1993 Jul;31(7):1927–1931. doi: 10.1128/jcm.31.7.1927-1931.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]