View More View Less
  • 1 Semmelweis University Institute of Medical Microbiology H-1089 Budapest Nagyvárad tér 4 Hungary
  • 2 University of Edinburgh Molecular Chemotherapy, Centre for Infectious Diseases 49 Little France Crescent Edinburgh EH16 4SB Scotland, UK
Restricted access

Purchase article

USD  $25.00

1 year subscription (Individual Only)

USD  $784.00

Recently, a PCR-derived method for serotyping Streptococcus pneumoniae has been devised to substitute the conventional antiserum phenotypic method. The method initially used a multiplex PCR reaction, dividing the isolates into 6 different groups based on the detected PCR gel pattern. In order to optimise and refine this crucial step, the Taguchi technique was employed, which can evaluate the individual effect of six parameters (in this case: primers, MgCl 2 , nucleotide mix, polymerase and buffer), with only 18 experiments; varying the parameter levels in an orthogonal matrix which suppresses the interactions between them. With this method, clear and sharp bands were observed in 5 experiments out of the 18, while the PCR did not work reliably in the remaining cases. In addition, the PCR-based technique could be rendered more economic by the 10-fold lowering of the quantities of two primers. The modified reaction yielded identical results to those obtained with the original method.Furthermore, we have designed serotype-specific primers for 11 new serotypes. The most important ones are those that can distinguish the very closely related, but equally important serotypes 6A and 6B.

  • WHO: Streptococcus pneumoniae (Pneumococcus). At http://www.who.int/immunization_delivery/new_vaccines/pneumo/en/

  • McGee, L., McDougal, L., Zhou, J., Spratt, B. G., Tenover, F. C., George, R., Hakenbeck, R., Hryniewicz, W., Lefévre, J. C., Tomasz, A., Klugman, K. P.: Nomenclature of major antimicrobial-resistant clones of Streptococcus pneumoniae defined by the Pneumococcal Molecular Epidemiology Network. J. Clin. Microbiol. 39 , 2565–2571 (2001).

    Klugman K. P. , 'Nomenclature of major antimicrobial-resistant clones of Streptococcus pneumoniae defined by the Pneumococcal Molecular Epidemiology Network ' (2001 ) 39 J. Clin. Microbiol. : 2565 -2571.

    • Search Google Scholar
  • de Lencastre, H., Tomasz, A.: From ecological reservoir to disease: the nasopharynx, day-care centres and drug-resistant clones of Streptococcus pneumoniae . J. Antimicrob. Chemother. 50 , 75–81 (2002).

    Tomasz A. , 'From ecological reservoir to disease: the nasopharynx, day-care centres and drug-resistant clones of Streptococcus pneumoniae ' (2002 ) 50 J. Antimicrob. Chemother. : 75 -81.

    • Search Google Scholar
  • Ledwith, M.: Pneumococcal conjugate vaccine. Curr. Opin. Pediatr. 13 , 70–74 (2001).

    Ledwith M. , 'Pneumococcal conjugate vaccine ' (2001 ) 13 Curr. Opin. Pediatr. : 70 -74.

  • Pletz, M. W., Maus, U., Krug, N., Welte, T., Lode, H.: Pneumococcal vaccines: mechanism of action, impact on epidemiology and adaptation of the species. Int. J. Antimicrob. Agents 32 , 199–206 (2008).

    Lode H. , 'Pneumococcal vaccines: mechanism of action, impact on epidemiology and adaptation of the species ' (2008 ) 32 Int. J. Antimicrob. Agents : 199 -206.

    • Search Google Scholar
  • Brito, D. A., Ramirez, M., de Lencastre, H.: Serotyping Streptococcus pneumoniae by multiplex PCR. J. Clin. Microbiol. 41 , 2378–2384 (2003).

    Lencastre H. , 'Serotyping Streptococcus pneumoniae by multiplex PCR ' (2003 ) 41 J. Clin. Microbiol. : 2378 -2384.

    • Search Google Scholar
  • Nagai, K., Shibasaki, Y., Hasegawa, K., Davies, T. A., Jacobs, M. R., Ubukata, K., Appelbaum, P. C.: Evaluation of PCR primers to screen for Streptococcus pneumoniae isolates and β -lactam resistance, and to detect common macrolide resistance determinants. J. Antimicrob. Chemother. 48 , 915–918 (2001).

    Appelbaum P. C. , 'Evaluation of PCR primers to screen for Streptococcus pneumoniae isolates and β-lactam resistance, and to detect common macrolide resistance determinants ' (2001 ) 48 J. Antimicrob. Chemother. : 915 -918.

    • Search Google Scholar
  • Taguchi, G.: Taguchi Methods . ASI Press, Dearborn, Michigan, 1993.

    Taguchi G. , '', in Taguchi Methods , (1993 ) -.

  • Jeney, Cs., Dobay, O., Lengyel, A., Ádám, É., Nász, I.: Taguchi optimisation of ELISA procedures. J. Immunol. Meth. 223 , 137–146 (1999).

    Nász I. , 'Taguchi optimisation of ELISA procedures ' (1999 ) 223 J. Immunol. Meth. : 137 -146.

    • Search Google Scholar
  • Clarke, S. C.: Control of pneumococcal disease in the United Kingdom — the start of a new era. J. Med. Microbiol. 55 , 975–980 (2006).

    Clarke S. C. , 'Control of pneumococcal disease in the United Kingdom — the start of a new era ' (2006 ) 55 J. Med. Microbiol. : 975 -980.

    • Search Google Scholar
  • EARSS Annual Report 2006. At http://www.rivm.nl/earss/result/Monitoring_reports/

  • Dobay, O., Rozgonyi, F., Hajdú, E., Nagy, E., Knausz, M., Amyes, S. G. B.: The relationship between serotypes and PFGE genotypes in isolates of Streptococcus pneumoniae from Hungary. Clin. Microb. Infect. 11 , 673–676 (2005).

    Amyes S. G. B. , 'The relationship between serotypes and PFGE genotypes in isolates of Streptococcus pneumoniae from Hungary ' (2005 ) 11 Clin. Microb. Infect. : 673 -676.

    • Search Google Scholar

Monthly Content Usage

Abstract Views Full Text Views PDF Downloads
Sep 2020 1 1 1
Oct 2020 5 0 0
Nov 2020 1 0 0
Dec 2020 0 0 0
Jan 2021 0 0 0
Feb 2021 3 0 0
Mar 2021 0 0 0