This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Brito, D. A.
Right arrow Articles by de Lencastre, H.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Brito, D. A.
Right arrow Articles by de Lencastre, H.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, June 2003, p. 2378-2384, Vol. 41, No. 6
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.6.2378-2384.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Serotyping Streptococcus pneumoniae by Multiplex PCR

D. A. Brito,1 M. Ramirez,1,2* and H. de Lencastre1,3

Laboratory of Molecular Genetics, Instituto de Tecnologia Química e Biológica da Universidade Nova de Lisboa, Oeiras,1 Laboratory of Microbiology, Faculdade de Medicina de Lisboa, Lisbon, Portugal,2 Laboratory of Microbiology, The Rockefeller University, New York, New York3

Received 16 December 2002/ Returned for modification 1 February 2003/ Accepted 18 February 2003

The capsule is a major virulence factor of pneumococci, and it was shown that some capsular variants are associated with antimicrobial resistance and certain types of disease. Moreover, pneumococcal capsular typing has received renewed interest since the availability of conjugate vaccines, which include serotypes frequently associated with pediatric disease. Our aim was to develop a simple, reliable, and economical method for detecting epidemiologically important serotypes present in the proposed 11-valent conjugate vaccine. We designed primers based on the sequences available for the capsular types 1, 3, 4, 6B, 14, 18C, 19F, 19A, and 23F and combined them into seven multiplex PCRs. The method involves streamlined DNA template preparation and agarose gel electrophoresis to analyze the amplification products. A total of 446 pneumococci selected from among isolates colonizing the nasopharynx of children attending day care centers in Lisbon, Portugal, were typed both by conventional immunological techniques and by multiplex PCR. Capsular types identified by the PCR method invariably produced results concordant with the conventional serotyping technique. Even when the method presented does not fully type an isolate, the PCR data can guide the experimenter when using immunological serotyping. Multiplex PCR for the analysis of pneumococci provides an accurate, expeditious, and cost-effective way of reducing the number of strains that have to be serotyped by conventional immunological techniques.

* Corresponding author. Mailing address: Laboratory of Molecular Genetics, Instituto de Tecnologia Química e Biológica, Rua Da Quinta Grande, n. 6, Apartado 127, 2780-156 Oeiras, Portugal. Phone: (351) 21-4469876. Fax: (351) 21-4428766. E-mail: ramirma{at}itqb.unl.pt.

Journal of Clinical Microbiology, June 2003, p. 2378-2384, Vol. 41, No. 6
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.6.2378-2384.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Jin, P., Xiao, M., Kong, F., Oftadeh, S., Zhou, F., Liu, C., Gilbert, G. L. (2009). Simple, Accurate, Serotype-Specific PCR Assay To Differentiate Streptococcus pneumoniae Serotypes 6A, 6B, and 6C. J. Clin. Microbiol. 47: 2470-2474 [Abstract] [Full Text]  
  • Thurmer, A., Helbig, J. H., Jacobs, E., Luck, P. C. (2009). PCR-based 'serotyping' of Legionella pneumophila. J Med Microbiol 58: 588-595 [Abstract] [Full Text]  
  • Nunes, S., Valente, C., Sa-Leao, R., de Lencastre, H. (2009). Temporal Trends and Molecular Epidemiology of Recently Described Serotype 6C of Streptococcus pneumoniae. J. Clin. Microbiol. 47: 472-474 [Abstract] [Full Text]  
  • Hotomi, M., Billal, D. S., Kamide, Y., Kanesada, K., Uno, Y., Kudo, F., Ito, M., Kakehata, S., Sugita, R., Ogami, M., Yamanaka, N., for the Advanced Treatment for Otitis Media Study, (2008). Serotype Distribution and Penicillin Resistance of Streptococcus pneumoniae Isolates from Middle Ear Fluids of Pediatric Patients with Acute Otitis Media in Japan. J. Clin. Microbiol. 46: 3808-3810 [Abstract] [Full Text]  
  • Yu, J., Carvalho, M. d. G. S., Beall, B., Nahm, M. H. (2008). A rapid pneumococcal serotyping system based on monoclonal antibodies and PCR. J Med Microbiol 57: 171-178 [Abstract] [Full Text]  
  • Nunes, S., Sa-Leao, R., de Lencastre, H. (2008). Optochin Resistance among Streptococcus pneumoniae Strains Colonizing Healthy Children in Portugal. J. Clin. Microbiol. 46: 321-324 [Abstract] [Full Text]  
  • Martins, E. R., Melo-Cristino, J., Ramirez, M. (2007). Reevaluating the Serotype II Capsular Locus of Streptococcus agalactiae. J. Clin. Microbiol. 45: 3384-3386 [Abstract] [Full Text]  
  • Dias, C. A., Teixeira, L. M., Carvalho, M. d. G., Beall, B. (2007). Sequential multiplex PCR for determining capsular serotypes of pneumococci recovered from Brazilian children. J Med Microbiol 56: 1185-1188 [Abstract] [Full Text]  
  • Szabo, J., Dobay, O., Erdos, M., Borbely, A., Rozgonyi, F., Marodi, L. (2007). Recurrent infection with genetically identical pneumococcal isolates in a patient with interleukin-1 receptor-associated kinase-4 deficiency. J Med Microbiol 56: 863-865 [Abstract] [Full Text]  
  • Izdebski, R., Sadowy, E., Fiett, J., Grzesiowski, P., Gniadkowski, M., Hryniewicz, W. (2007). Clonal Diversity and Resistance Mechanisms in Tetracycline-Nonsusceptible Streptococcus pneumoniae Isolates in Poland. Antimicrob. Agents Chemother. 51: 1155-1163 [Abstract] [Full Text]  
  • Carrico, J. A., Silva-Costa, C., Melo-Cristino, J., Pinto, F. R., de Lencastre, H., Almeida, J. S., Ramirez, M. (2006). Illustration of a Common Framework for Relating Multiple Typing Methods by Application to Macrolide-Resistant Streptococcus pyogenes.. J. Clin. Microbiol. 44: 2524-2532 [Abstract] [Full Text]  
  • Lin, J., Kaltoft, M. S., Brandao, A. P., Echaniz-Aviles, G., Brandileone, M. C. C., Hollingshead, S. K., Benjamin, W. H., Nahm, M. H. (2006). Validation of a Multiplex Pneumococcal Serotyping Assay with Clinical Samples. J. Clin. Microbiol. 44: 383-388 [Abstract] [Full Text]  
  • Sa-Leao, R., Simoes, A. S., Nunes, S., Sousa, N. G., Frazao, N., de Lencastre, H. (2006). Identification, prevalence and population structure of non-typable Streptococcus pneumoniae in carriage samples isolated from preschoolers attending day-care centres. Microbiology 152: 367-376 [Abstract] [Full Text]  
  • Pai, R., Gertz, R. E., Beall, B. (2006). Sequential Multiplex PCR Approach for Determining Capsular Serotypes of Streptococcus pneumoniae Isolates. J. Clin. Microbiol. 44: 124-131 [Abstract] [Full Text]  
  • Moreno, J., Hernandez, E., Sanabria, O., Castaneda, E. (2005). Detection and Serotyping of Streptococcus pneumoniae from Nasopharyngeal Samples by PCR-Based Multiplex Assay. J. Clin. Microbiol. 43: 6152-6154 [Abstract] [Full Text]  
  • Pai, R., Limor, J., Beall, B. (2005). Use of Pyrosequencing To Differentiate Streptococcus pneumoniae Serotypes 6A and 6B. J. Clin. Microbiol. 43: 4820-4822 [Abstract] [Full Text]  
  • Tonnaer, E. L., Rijkers, G. T., Meis, J. F., Klaassen, C. H., Bogaert, D., Hermans, P. W., Curfs, J. H. (2005). Genetic Relatedness between Pneumococcal Populations Originating from the Nasopharynx, Adenoid, and Tympanic Cavity of Children with Otitis Media. J. Clin. Microbiol. 43: 3140-3144 [Abstract] [Full Text]  
  • O'Halloran, D. M., Cafferkey, M. T. (2005). Multiplex PCR for Identification of Seven Streptococcus pneumoniae Serotypes Targeted by a 7-Valent Conjugate Vaccine. J. Clin. Microbiol. 43: 3487-3490 [Abstract] [Full Text]  
  • Batt, S. L., Charalambous, B. M., McHugh, T. D., Martin, S., Gillespie, S. H. (2005). Novel PCR-Restriction Fragment Length Polymorphism Method for Determining Serotypes or Serogroups of Streptococcus pneumoniae Isolates. J. Clin. Microbiol. 43: 2656-2661 [Abstract] [Full Text]  
  • Yershov, A. L., Jordan, B. S., Guymon, C. H., Dubick, M. A. (2005). Relationship between the inoculum dose of Streptococcus pneumoniae and pneumonia onset in a rabbit model. Eur Respir J 25: 693-700 [Abstract] [Full Text]  
  • Birtles, A., McCarthy, N., Sheppard, C. L., Rutter, H., Guiver, M., Haworth, E., George, R. C. (2005). Multilocus Sequence Typing Directly on DNA from Clinical Samples and a Cultured Isolate To Investigate Linked Fatal Pneumococcal Disease in Residents of a Shelter for Homeless Men. J. Clin. Microbiol. 43: 2004-2008 [Abstract] [Full Text]  
  • Yu, J., Lin, J., Benjamin, W. H. Jr., Waites, K. B., Lee, C.-h., Nahm, M. H. (2005). Rapid Multiplex Assay for Serotyping Pneumococci with Monoclonal and Polyclonal Antibodies. J. Clin. Microbiol. 43: 156-162 [Abstract] [Full Text]  
  • Rubin, L. G., Rizvi, A. (2004). PCR-based assays for detection of Streptococcus pneumoniae serotypes 3, 14, 19F and 23F in respiratory specimens. J Med Microbiol 53: 595-602 [Abstract] [Full Text]  
  • Slotved, H.-C., Kaltoft, M., Skovsted, I. C., Kerrn, M. B., Espersen, F. (2004). Simple, Rapid Latex Agglutination Test for Serotyping of Pneumococci (Pneumotest-Latex). J. Clin. Microbiol. 42: 2518-2522 [Abstract] [Full Text]  
  • Kong, F., Gilbert, G. L. (2003). Using cpsA-cpsB sequence polymorphisms and serotype-/group-specific PCR to predict 51 Streptococcus pneumoniae capsular serotypes. J Med Microbiol 52: 1047-1058 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»