Advertisement

Unravelling the Ecology of Antibiotic Resistant Bacteria in the Nasopharynx

  • Bambos M. CharalambousEmail author
  • Ndeky M. Oriyo
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 642)

Abstract

To study the dynamics and diversity of pneumococcal carriage and antibiotic resistance, a more thorough and systematic approach has been employed compared with routine surveillance of serotype and anti­biotic resistance. Up to ten pneumococcal isolates from pernasal (nose) and oropharyngeal (throat) sites are isolated and characterised. Our carriage studies have revealed a diverse community of pneumococci with multiple strains colonising the nasopharynx of children. In Tanzanian children less than 6 years of age, up to six serotypes and up to six different antibiotic sensitivities (as distinguished by at least a fourfold difference in the minimum inhibitory concentration) have been found. Serotyping by the Quelling reaction is prone to inaccuracy and requires expensive serological reagents. To improve the accuracy and reduce the costs, an alternative capsular typing DNA-based method has been developed. This chapter will describe the methods we have employed with emphasis on the capsular typing method.

Key words

Streptococcus pneumoniae Capsular typing Nasopharyngeal colonisation Antibiotic resistance Diversity 

Notes

Acknowledgments

The authors wish to thank Dr. Steve Platt for his developing the novel Bionumerics script for the cluster analysis of in vitro and in silico data. We also thank Dr. Claire Jenkins for the data analysis shown in Figs. 2 and 3.

References

  1. 1.
    Gray BM, Converse GM III, Dillon HC (1980) J Infect Dis 142:923–933PubMedGoogle Scholar
  2. 2.
    Faden H, Duffy L, Wasielewski R, Wolf J, Krystofik D, Tung Y (1997) J Infect Dis 175:1440–1445CrossRefPubMedGoogle Scholar
  3. 3.
    Stromberg A, Friberg U, Cars O (1987) Eur J Clin Microbiol 6:525–529CrossRefPubMedGoogle Scholar
  4. 4.
    Lipsitch M (2001) Am J Epidemiol 154:85–92CrossRefPubMedGoogle Scholar
  5. 5.
    Coffey TJ, Dowson CG, Daniels M, Zhou J, Martin C, Spratt BG, Musser JM (1991) Mol Microbiol 5:2255–2260CrossRefPubMedGoogle Scholar
  6. 6.
    Coffey TJ, Enright MC, Daniels M, Morona JK, Morona R, Hryniewicz W, Paton JC, Spratt BG (1998) Mol Microbiol 27:73–83CrossRefPubMedGoogle Scholar
  7. 7.
    Dowson CG, Hutchison A, Brannigan JA, George RC, Hansman D, Liñares J, Tomasz A, Smith JM, Spratt BG (1989) Proc Natl Acad Sci USA 86:8842–8846CrossRefPubMedGoogle Scholar
  8. 8.
    Feil EJ, Enright MC, Spratt BG (2000) Res Microbiol 151:465–469CrossRefPubMedGoogle Scholar
  9. 9.
    Jefferies JM, Smith A, Clarke SC, Dowson C, Mitchell TJ (2004) J Clin Microbiol 42:5681–5688CrossRefPubMedGoogle Scholar
  10. 10.
    Charalambous BM, Oriyo NM, Gillespie SH (2008) J Clin Microbiol 46:2467–2468CrossRefPubMedGoogle Scholar
  11. 11.
    Hiller NL, Janto B, Hogg JS, Boissy R, Yu S, Powell E, Keefe R, Ehrlich NE, Shen K, Hayes J, Barbadora K, Klimke W, Dernovoy D, Tatusova T, Parkhill J, Bentley SD, Post JC, Ehrlich GD, Hu FZ (2007) J Bacteriol 189:8186–8195CrossRefPubMedGoogle Scholar
  12. 12.
    Whatmore AM, Barcus VA, Dowson CG (1999) J Bacteriol 181:3144–3154PubMedGoogle Scholar
  13. 13.
    Poulsen K, Reinholdt J, Jespersgaard C, Boye K, Brown TA, Hauge M, Kilian M (1998) Infect Immun 66:181–190PubMedGoogle Scholar
  14. 14.
    Gundel M, Okura G (1933) Z Hyg Infect 114:678–704CrossRefGoogle Scholar
  15. 15.
    Gratten M, Manning K, Dixon J, Morey F, Torzillo P, Hanna J, Erlich J, Asche V, Riley I (1994) Southeast Asian J Trop Med Public Health 25:123–131PubMedGoogle Scholar
  16. 16.
    Gratten M, Montgomery J, Gerega G, Gratten H, Siwi H, Poli A, Koki G (1989) Southeast Asian J Trop Med Public Health 20:501–509PubMedGoogle Scholar
  17. 17.
    Hansman D, Morris S, Gregory M, McDonald B (1985) J Hyg (Lond) 95:677–684CrossRefGoogle Scholar
  18. 18.
    Hansman D, Morris S (1988) Epidemiol Infect 101:411–417CrossRefPubMedGoogle Scholar
  19. 19.
    Billal DS, Hotomi M, Suzumoto M, Yamauchi K, Arai J, Katsurahara T, Moriyama S, Fujihara K, Yamanaka N (2008) Eur J Pediatr 167:401–407CrossRefPubMedGoogle Scholar
  20. 20.
    Pai R, Gertz RE, Beall B (2006) J Clin Microbiol 44:124–131CrossRefPubMedGoogle Scholar
  21. 21.
    Lawrence ER, Griffiths DB, Martin SA, George RC, Hall LM (2003) J Clin Microbiol 41:601–607CrossRefPubMedGoogle Scholar
  22. 22.
    Lawrence ER, Arias CA, Duke B, Beste D, Broughton K, Efstratiou A, George RC, Hall LM (2000) J Clin Microbiol 38:1319–1323PubMedGoogle Scholar
  23. 23.
    Batt SL, Charalambous BM, McHugh TD, Martin S, Gillespie SH (2005) J Clin Microbiol 43:2656–2661CrossRefPubMedGoogle Scholar
  24. 24.
    Arnold C, Metherell L, Clewley JP, Stanley J (1999) Res Microbiol 150:33–44CrossRefPubMedGoogle Scholar
  25. 25.
    Wei W, Davis RE, Lee IM, Zhao Y (2007) Int J Syst Evol Microbiol 57:1855–1867CrossRefPubMedGoogle Scholar
  26. 26.
    Sims EJ, Goyal M, Arnold C (2002) J Clin Microbiol 40:4072–4076CrossRefPubMedGoogle Scholar
  27. 27.
    Dice LR (1945) Ecology 26:297–302CrossRefGoogle Scholar
  28. 28.
    Sneath PH, Sokal RR (1973) Numerical taxonomy: the principles and practice of numerical classification. W.H. Freeman, San FranciscoGoogle Scholar
  29. 29.
    Charalambous BM, Batt SL, Peek AC, Mwerinde H, Sam N, Gillespie SH (2003) J Clin Microbiol 41:5551–5556CrossRefPubMedGoogle Scholar
  30. 30.
    Bauer AW, Kirby WM, Sherris JC, Turck M (1966) Tech Bull Regist Med Technol 36:49–52PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  1. 1.Centre for Clinical Microbiology Department of InfectionUniversity College LondonLondonUK
  2. 2.Clinical LaboratoryKilimanjaro Christian Medical Centre, Tumaini UniversityMoshiTanzania

Personalised recommendations