Korean J Pediatr.  2012 Nov;55(11):424-429. 10.3345/kjp.2012.55.11.424.

Enhanced detection and serotyping of Streptococcus pneumoniae using multiplex polymerase chain reaction

• Ahn JG
• Choi SY
• Kim DS
• Kim KH

• KMID: 1781704
• DOI: http://doi.org/10.3345/kjp.2012.55.11.424

Abstract

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 Children's 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.

Keyword

Streptococcus pneumoniae; Multiplex polymerase chain reaction; Culture

MeSH Terms

Culture Techniques
Humans
Korea
Multiplex Polymerase Chain Reaction
Pneumonia
Polymerase Chain Reaction
Respiratory Tract Infections
Serotyping
Streptococcus
Streptococcus pneumoniae