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Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1995 Jun;33(6):1606–1612. doi: 10.1128/jcm.33.6.1606-1612.1995

Comparative study of five different DNA fingerprint techniques for molecular typing of Streptococcus pneumoniae strains.

P W Hermans 1, M Sluijter 1, T Hoogenboezem 1, H Heersma 1, A van Belkum 1, R de Groot 1
PMCID: PMC228225  PMID: 7650196

Abstract

The aim of this study was to identify the strengths and weaknesses of five DNA fingerprint methods for epidemiological typing of Streptococcus pneumoniae. We investigated the usefulness of (i) ribotyping, (ii) BOX fingerprinting with the BOX repetitive sequence of S. pneumoniae as a DNA probe, (iii) PCR fingerprinting with a primer homologous to the enterobacterial repetitive intergenic consensus sequence, (iv) pulsed-field gel electrophoresis of large DNA fragments, and (v) restriction fragment end labeling to detect restriction fragment length polymorphism of small DNA fragments. Twenty-eight S. pneumoniae strains isolated from the blood and/or cerebrospinal fluid of 21 patients were analyzed. Genetic clustering among the 28 strains was independent of the DNA fingerprint technique used. However, the discriminatory power and the similarity values differed significantly among the individual techniques. BOX fingerprinting, pulsed-field gel electrophoresis, and restriction fragment end labeling provided the highest degree of discriminatory power. Furthermore, the ease with which computerized fingerprint analysis could be conducted also varied significantly among the techniques. Ribotyping, BOX fingerprinting, and restriction fragment end labeling were very suitable techniques for accurate computerized data analysis. Because of their high discriminatory potential and ease of accurate analysis, we conclude that BOX fingerprinting and restriction fragment end labeling are the most suitable techniques to type pneumococcal strains.

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Selected References

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  1. Cherian T., Steinhoff M. C., Harrison L. H., Rohn D., McDougal L. K., Dick J. A cluster of invasive pneumococcal disease in young children in child care. JAMA. 1994 Mar 2;271(9):695–697. [PubMed] [Google Scholar]
  2. Dowson C. G., Hutchison A., Brannigan J. A., George R. C., Hansman D., Liñares J., Tomasz A., Smith J. M., Spratt B. G. Horizontal transfer of penicillin-binding protein genes in penicillin-resistant clinical isolates of Streptococcus pneumoniae. Proc Natl Acad Sci U S A. 1989 Nov;86(22):8842–8846. doi: 10.1073/pnas.86.22.8842. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Fenoll A., Martín Bourgon C., Muñz R., Vicioso D., Casal J. Serotype distribution and antimicrobial resistance of Streptococcus pneumoniae isolates causing systemic infections in Spain, 1979-1989. Rev Infect Dis. 1991 Jan-Feb;13(1):56–60. doi: 10.1093/clinids/13.1.56. [DOI] [PubMed] [Google Scholar]
  4. Hermans P. W., van Soolingen D., Bik E. M., de Haas P. E., Dale J. W., van Embden J. D. Insertion element IS987 from Mycobacterium bovis BCG is located in a hot-spot integration region for insertion elements in Mycobacterium tuberculosis complex strains. Infect Immun. 1991 Aug;59(8):2695–2705. doi: 10.1128/iai.59.8.2695-2705.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Kell C. M., Jordens J. Z., Daniels M., Coffey T. J., Bates J., Paul J., Gilks C., Spratt B. G. Molecular epidemiology of penicillin-resistant pneumococci isolated in Nairobi, Kenya. Infect Immun. 1993 Oct;61(10):4382–4391. doi: 10.1128/iai.61.10.4382-4391.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Kristinsson K. G., Hjálmarsdóttir M. A., Steingrímsson O. Increasing penicillin resistance in pneumococci in Iceland. Lancet. 1992 Jun 27;339(8809):1606–1607. doi: 10.1016/0140-6736(92)91868-9. [DOI] [PubMed] [Google Scholar]
  7. Lefevre J. C., Faucon G., Sicard A. M., Gasc A. M. DNA fingerprinting of Streptococcus pneumoniae strains by pulsed-field gel electrophoresis. J Clin Microbiol. 1993 Oct;31(10):2724–2728. doi: 10.1128/jcm.31.10.2724-2728.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Martin B., Humbert O., Camara M., Guenzi E., Walker J., Mitchell T., Andrew P., Prudhomme M., Alloing G., Hakenbeck R. A highly conserved repeated DNA element located in the chromosome of Streptococcus pneumoniae. Nucleic Acids Res. 1992 Jul 11;20(13):3479–3483. doi: 10.1093/nar/20.13.3479. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Marton A., Gulyas M., Munoz R., Tomasz A. Extremely high incidence of antibiotic resistance in clinical isolates of Streptococcus pneumoniae in Hungary. J Infect Dis. 1991 Mar;163(3):542–548. doi: 10.1093/infdis/163.3.542. [DOI] [PubMed] [Google Scholar]
  10. McDougal L. K., Facklam R., Reeves M., Hunter S., Swenson J. M., Hill B. C., Tenover F. C. Analysis of multiply antimicrobial-resistant isolates of Streptococcus pneumoniae from the United States. Antimicrob Agents Chemother. 1992 Oct;36(10):2176–2184. doi: 10.1128/aac.36.10.2176. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Munoz R., Musser J. M., Crain M., Briles D. E., Marton A., Parkinson A. J., Sorensen U., Tomasz A. Geographic distribution of penicillin-resistant clones of Streptococcus pneumoniae: characterization by penicillin-binding protein profile, surface protein A typing, and multilocus enzyme analysis. Clin Infect Dis. 1992 Jul;15(1):112–118. doi: 10.1093/clinids/15.1.112. [DOI] [PubMed] [Google Scholar]
  12. Muñoz R., Coffey T. J., Daniels M., Dowson C. G., Laible G., Casal J., Hakenbeck R., Jacobs M., Musser J. M., Spratt B. G. Intercontinental spread of a multiresistant clone of serotype 23F Streptococcus pneumoniae. J Infect Dis. 1991 Aug;164(2):302–306. doi: 10.1093/infdis/164.2.302. [DOI] [PubMed] [Google Scholar]
  13. Muñoz R., Dowson C. G., Daniels M., Coffey T. J., Martin C., Hakenbeck R., Spratt B. G. Genetics of resistance to third-generation cephalosporins in clinical isolates of Streptococcus pneumoniae. Mol Microbiol. 1992 Sep;6(17):2461–2465. doi: 10.1111/j.1365-2958.1992.tb01422.x. [DOI] [PubMed] [Google Scholar]
  14. Nielsen S. V., Henrichsen J. Capsular types and susceptibility to penicillin of pneumococci isolated from cerebrospinal fluid or blood in Denmark, 1983-1988. Scand J Infect Dis. 1993;25(2):165–170. doi: 10.3109/00365549309008480. [DOI] [PubMed] [Google Scholar]
  15. Sessegolo J. F., Levin A. S., Levy C. E., Asensi M., Facklam R. R., Teixeira L. M. Distribution of serotypes and antimicrobial resistance of Streptococcus pneumoniae strains isolated in Brazil from 1988 to 1992. J Clin Microbiol. 1994 Apr;32(4):906–911. doi: 10.1128/jcm.32.4.906-911.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Shapiro E. D., Austrian R. Serotypes responsible for invasive Streptococcus pneumoniae infections among children in Connecticut. J Infect Dis. 1994 Jan;169(1):212–214. doi: 10.1093/infdis/169.1.212. [DOI] [PubMed] [Google Scholar]
  17. Smith A. M., Klugman K. P., Coffey T. J., Spratt B. G. Genetic diversity of penicillin-binding protein 2B and 2X genes from Streptococcus pneumoniae in South Africa. Antimicrob Agents Chemother. 1993 Sep;37(9):1938–1944. doi: 10.1128/aac.37.9.1938. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Soares S., Kristinsson K. G., Musser J. M., Tomasz A. Evidence for the introduction of a multiresistant clone of serotype 6B Streptococcus pneumoniae from Spain to Iceland in the late 1980s. J Infect Dis. 1993 Jul;168(1):158–163. doi: 10.1093/infdis/168.1.158. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. van Belkum A. DNA fingerprinting of medically important microorganisms by use of PCR. Clin Microbiol Rev. 1994 Apr;7(2):174–184. doi: 10.1128/cmr.7.2.174. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. van Soolingen D., de Haas P. E., Hermans P. W., van Embden J. D. DNA fingerprinting of Mycobacterium tuberculosis. Methods Enzymol. 1994;235:196–205. doi: 10.1016/0076-6879(94)35141-4. [DOI] [PubMed] [Google Scholar]

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