Advanced Search
Korean J Pediatr 2012 November;55(11) :424-429.
Published online 2012 July 23.        doi:
Enhanced detection and serotyping of Streptococcus pneumoniae using multiplex polymerase chain reaction
Jong Gyun Ahn (Ahn JG)1,2, Seong Yeol Choi (Choi SY)2, Dong Soo Kim (Kim DS)2,3, Ki Hwan Kim (Kim KH)2,3
1,2Department of Pediatrics, School of Medicine, Ewha Womans University, Seoul, Korea
2,3Department of Pediatrics, Severance Childrens Hospital, Yonsei University College of Medicine, Seoul, Korea
2Department of Medicine, The Graduate School of Yonsei University, Seoul, Korea
Corresponding Author: Ki Hwan Kim ,Tel: +82-2-2228-2050, Fax: +82-2-393-9118, Email:
Copyright © 2012 by The Korean Pediatric Society
Purpose: Methods for quick and reliable detection of Streptococcus pneumoniae are needed for the diagnosis of pneumococcal disease and vaccine studies. This study aimed to show that sequential multiplex polymerase chain reaction (PCR) is more efficient than conventional culture in achieving S. pneumoniae -positive results. Methods: Nasopharyngeal (NP) secretions were obtained from 842 pediatric patients admitted with lower respiratory infections at Severance Childrens Hospital in Korea between March 2009 and June 2010. For identification and serotype determination of pneumococci from the NP secretions, the secretions were evaluated via multiplex PCR technique with 35 serotype-specific primers arranged in 8 multiplex PCR sets and conventional bacteriological culture technique. Results: Among the results for 793 samples that underwent both bacterial culture and PCR analysis for pneumococcal detection, 153 (19.3%) results obtained by PCR and 81 (10.2%) results obtained by conventional culture technique were positive for S. pneumoniae. The predominant serotypes observed, in order of decreasing frequency, were 19A (23%), 6A/B (16%), 19F (11%), 15B/C (5%), 15A (5%), and 11A (4%); further, 26% of the isolates were non-typeable. Conclusion: As opposed to conventional bacteriological tests, PCR analysis can accurately and rapidly identify pneumococcal serotypes.
Keywords: Streptococcus pneumoniae | Multiplex polymerase chain reaction | Culture
1. Pneumococcal conjugate vaccine for childhood immunization: WHO position paper. Wkly Epidemiol Rec 2007;82:93–104.
2. Hausdorff WP, Bryant J, Paradiso PR, Siber GR. Which pneumococcal serogroups cause the most invasive disease: implications for conjugate vaccine formulation and use, part I. Clin Infect Dis 2000;30:100–121.
3. Millar EV, O'Brien KL, Watt JP, Bronsdon MA, Dallas J, Whitney CG, et al. Effect of community-wide conjugate pneumococcal vaccine use in infancy on nasopharyngeal carriage through 3 years of age: a cross-sectional study in a high-risk population. Clin Infect Dis 2006;43:8–15.
4. Hanage WP, Huang SS, Lipsitch M, Bishop CJ, Godoy D, Pelton SI, et al. Diversity and antibiotic resistance among nonvaccine serotypes of Streptococcus pneumoniae carriage isolates in the post-heptavalent conjugate vaccine era. J Infect Dis 2007;195:347–352.
5. Tan TQ. Serious and invasive pediatric pneumococcal disease: epidemiology and vaccine impact in the USA. Expert Rev Anti Infect Ther 2010;8:117–12
6. Isaacman DJ, McIntosh ED, Reinert RR. Burden of invasive pneumococcal disease and serotype distribution among Streptococcus pneumoniae isolates in young children in Europe: impact of the 7-valent pneumococcal conjugate vaccine and considerations for future conjugate vaccines. Int J Infect Dis 2010;14:e197–e209.
7. Lin TY, Shah NK, Brooks D, Garcia CS. Summary of invasive pneumococcal disease burden among children in the Asia-Pacific region. Vaccine 2010;28:7589–7605.
8. Whitney CG, Pilishvili T, Farley MM, Schaffner W, Craig AS, Lynfield R, et al. Effectiveness of seven-valent pneumococcal conjugate vaccine against invasive pneumococcal disease: a matched case-control study. Lancet 2006;368:1495–1502.
9. Centers for Disease Control and Prevention (CDC). Invasive pneumococcal disease in children 5 years after conjugate vaccine introduction--eight states, 1998-2005. MMWR Morb Mortal Wkly Rep 2008;57:144–148.
10. Huang SS, Hinrichsen VL, Stevenson AE, Rifas-Shiman SL, Kleinman K, Pelton SI, et al. Continued impact of pneumococcal conjugate vaccine on carriage in young children. Pediatrics 2009;124:e1–e11.
11. Ghaffar F, Barton T, Lozano J, Muniz LS, Hicks P, Gan V, et al. Effect of the 7-valent pneumococcal conjugate vaccine on nasopharyngeal colonization by Streptococcus pneumoniae in the first 2 years of life. Clin Infect Dis 2004;39:930–938.
12. Moore MR. Rethinking replacement and resistance. J Infect Dis 2009;199:771–773.
13. Dagan R, Givon-Lavi N, Leibovitz E, Greenberg D, Porat N. Introduction and proliferation of multidrug-resistant Streptococcus pneumoniae serotype 19A clones that cause acute otitis media in an unvaccinated population. J Infect Dis 2009;199:776–785.
14. Pai R, Gertz RE, Beall B. Sequential multiplex PCR approach for determining capsular serotypes of Streptococcus pneumoniae isolates. J Clin Microbiol 2006;44:124–131.
15. Bentley SD, Aanensen DM, Mavroidi A, Saunders D, Rabbinowitsch E, Collins M, et al. Genetic analysis of the capsular biosynthetic locus from all 90 pneumococcal serotypes. PLoS Genet 2006;2:e31.
16. Saha SK, Darmstadt GL, Baqui AH, Hossain B, Islam M, Foster D, et al. Identification of serotype in culture negative pneumococcal meningitis using sequential multiplex PCR: implication for surveillance and vaccine design. PLoS One 2008;3:e3576.
17. Lee TJ, Chun JK, Kim KH, Kim KJ, Kim DS. Serotype distribution of pneumococcus isolated from the ear discharge in children with otitis media in 2001-2006. Korean J Pediatr Infect Dis 2008;15:44–50.
18. Song JH, Baek JY, Cheong HS, Chung DR, Peck KR, Ko KS. Changes of serotype and genotype in Streptococcus pneumoniae isolates from a Korean hospital in 2007. Diagn Microbiol Infect Dis 2009;63:271–278.
19. Choi EH, Kim SH, Eun BW, Kim SJ, Kim NH, Lee J, et al. Streptococcus pneumoniae serotype 19A in children, South Korea. Emerg Infect Dis 2008;14:275–281.
20. Virolainen A, Salo P, Jero J, Karma P, Eskola J, Leinonen M. Comparison of PCR assay with bacterial culture for detecting Streptococcus pneumoniae in middle ear fluid of children with acute otitis media. J Clin Microbiol 1994;32:2667–2670.
21. Kearns AM, Freeman R, Murphy OM, Seiders PR, Steward M, Wheeler J. Rapid PCR-based detection of Streptococcus pneumoniae DNA in cerebrospinal fluid. J Clin Microbiol 1999;37:3434.
22. Kontiokari T, Renko M, Kaijalainen T, Kuisma L, Leinonen M. Comparison of nasal swab culture, quantitative culture of nasal mucosal tissue and PCR in detecting Streptococcus pneumoniae carriage in rats. APMIS 2000;108:734–738.
23. Brugger SD, Hathaway LJ, Muhlemann K. Detection of Streptococcus pneumoniae strain cocolonization in the nasopharynx. J Clin Microbiol 2009;47:1750–1756.
24. O'Brien KL, Nohynek H. World Health Organization Pneumococcal Vaccine Trials Carriage Working Group. Report from a WHO Working Group: standard method for detecting upper respiratory carriage of Streptococcus pneumoniae. Pediatr Infect Dis J 2003;22:e1–e11.
25. Arai S, Konda T, Wad A, Matsunaga Y, Okabe N, Watanabe H, et al. Use of antiserum-coated latex particles for serotyping Streptococcus pneumoniae. Microbiol Immunol 2001;45:159–162.
26. Morais L, Carvalho Mda G, Roca A, Flannery B, Mandomando I, Soriano-Gabarro M, et al. Sequential multiplex PCR for identifying pneumococcal capsular serotypes from South-Saharan African clinical isolates. J Med Microbiol 2007;56(Pt 9):1181–1184.
27. Dias CA, Teixeira LM, Carvalho Mda G, Beall B. Sequential multiplex PCR for determining capsular serotypes of pneumococci recovered from Brazilian children. J Med Microbiol 2007;56(Pt 9):1185–1188.
28. Kim SM, Hur JK, Lee KY, Shin YK, Park SE, Ma SH, et al. Epidemiological study of pneumococcal nasal carriage and serotypes among Korean children. Korean J Pediatr 2004;47:611–616.
29. Korea Centers for Disease Control and Prevention. Serotype distribution and antimicrobial susceptibility of invasive Streptococcus pneumoniae clinical isolates in Korea. Public Health Wkly Rep 2010;3:1–7.
PDF Links  PDF Links
Full text via DOI  Full text via DOI
  via Pubmed
Full text via PMC  Full text via PMC
via Pubreader  via PubReader
Download Citation  Download Citation
Supplementary Material  Supplementary Material
A study of serotyping of Streptococcus pneumoniae by multibead assay  2007 February;50(2)
Pneumococcal Surface Protein A of Streptococcus pneumoniae Isolates from Koreans  2005 November;48(11)
Detection of Mycoplasma pneumoniae in clinical Samples from Pediatric Patients by Polymerase Chain Reaction  1995 August;38(8)
Detection of Cytomegalovirus DNA in Urine Culture Using Polymerase Chain Reaction  1993 May;36(5)
Register for e-submission
Register here to access the e-submission system of Korean J Pediatr for authors and reviewers.
Manuscript Submission
To submit a manuscript, please visit the Korean J Pediatr e-submission management system at, read the Instructions for Authors, and log into the Korean J Pediatr e-submission system. For assistance with manuscript submission, please contact:
Free archive
Anyone may access any past or current articles without logging in.
Korean Pediatric Society Office
#1606, Seocho World Officetel, 19 Seoun-ro, Seocho-gu, Seoul 137-070, Korea
TEL : +82-2-3473-7305    FAX : +82-2-3473-7307   E-mail:
BrowseCurrent IssueFor Authors and ReviewersAbout
Copyright© The Korean Pediatric Society. All right reserved.