Evaluation of semiautomated multiplex PCR assay for determination of Streptococcus pneumoniae serotypes and serogroups
- PMID: 12574253
- PMCID: PMC149661
- DOI: 10.1128/JCM.41.2.601-607.2003
Evaluation of semiautomated multiplex PCR assay for determination of Streptococcus pneumoniae serotypes and serogroups
Abstract
A semiautomated method for the determination of five serotypes and three serogroups in Streptococcus pneumoniae was developed. Primers specific for serotypes 1, 3, 14, 19F, and 23F and serogroups 6, 19, and 23 were combined in three multiplex PCRs. Products were separated by capillary electrophoresis with a 7-min run time, and a serotype or serogroup was assigned on the basis of fragment size. The method was used to test 93 clinical isolates, and all isolates of the serotypes concerned were correctly detected. The strategy would allow the detection of multiple serotypes in a single sample. Detection of additional serotypes could be included as capsule locus sequences become available.
Figures

Similar articles
-
Practical Prediction of Ten Common Streptococcus pneumoniae Serotypes/Serogroups in One PCR Reaction by Multiplex Ligation-Dependent Probe Amplification and Melting Curve (MLPA-MC) Assay in Shenzhen, China.PLoS One. 2015 Jul 7;10(7):e0130664. doi: 10.1371/journal.pone.0130664. eCollection 2015. PLoS One. 2015. PMID: 26151828 Free PMC article.
-
PCR-based assays for detection of Streptococcus pneumoniae serotypes 3, 14, 19F and 23F in respiratory specimens.J Med Microbiol. 2004 Jul;53(Pt 7):595-602. doi: 10.1099/jmm.0.45550-0. J Med Microbiol. 2004. PMID: 15184528
-
PCR-Based Serotyping of Streptococcus pneumoniae from Culture-Negative Specimens: Novel Primers for Detection of Serotypes within Serogroup 18.J Clin Microbiol. 2016 Aug;54(8):2178-81. doi: 10.1128/JCM.00419-16. Epub 2016 Jun 1. J Clin Microbiol. 2016. PMID: 27252464 Free PMC article.
-
Novel PCR-restriction fragment length polymorphism method for determining serotypes or serogroups of Streptococcus pneumoniae isolates.J Clin Microbiol. 2005 Jun;43(6):2656-61. doi: 10.1128/JCM.43.6.2656-2661.2005. J Clin Microbiol. 2005. PMID: 15956380 Free PMC article.
-
Current challenges in the accurate identification of Streptococcus pneumoniae and its serogroups/serotypes in the vaccine era.J Microbiol Methods. 2017 Oct;141:48-54. doi: 10.1016/j.mimet.2017.07.015. Epub 2017 Aug 2. J Microbiol Methods. 2017. PMID: 28780272 Review.
Cited by
-
Scalable transcriptional analysis routine--multiplexed quantitative real-time polymerase chain reaction platform for gene expression analysis and molecular diagnostics.J Mol Diagn. 2005 Oct;7(4):444-54. doi: 10.1016/S1525-1578(10)60575-2. J Mol Diagn. 2005. PMID: 16237214 Free PMC article.
-
Detection and serotyping of Streptococcus pneumoniae from nasopharyngeal samples by PCR-based multiplex assay.J Clin Microbiol. 2005 Dec;43(12):6152-4. doi: 10.1128/JCM.43.12.6152-6154.2005. J Clin Microbiol. 2005. PMID: 16333118 Free PMC article.
-
Evaluation of pneumococcal serotyping by multiplex PCR and quellung reactions.J Clin Microbiol. 2013 Dec;51(12):4193-5. doi: 10.1128/JCM.01876-13. Epub 2013 Sep 11. J Clin Microbiol. 2013. PMID: 24025905 Free PMC article.
-
Molecular characterization and antimicrobial susceptibility of Streptococcus pneumoniae isolated from children hospitalized with respiratory infections in Suzhou, China.PLoS One. 2014 Apr 7;9(4):e93752. doi: 10.1371/journal.pone.0093752. eCollection 2014. PLoS One. 2014. PMID: 24710108 Free PMC article. Clinical Trial.
-
Single-Step Multiplex PCR Assay for Determining 92 Pneumococcal Serotypes.J Clin Microbiol. 2016 Aug;54(8):2197-200. doi: 10.1128/JCM.01156-16. Epub 2016 Jun 8. J Clin Microbiol. 2016. PMID: 27280423 Free PMC article.
References
-
- Altschul, S. F., W. Gish, W. Miller, E. W. Myers, and D. J. Lipman. 1990. Basic local alignment search tool. J. Mol. Biol. 215:403-410. - PubMed
-
- Arrecubieta, C., E. Garcia, and R. Lopez. 1995. Sequence and transcriptional analysis of a DNA region involved in the production of capsular polysaccharide in Streptococcus pneumoniae type 3. Gene 167:1-7. - PubMed
-
- Ballagi-Pordany, A., and S. Belak. 1996. The use of mimics as internal standards to avoid false negatives in diagnostic PCR. Mol. Cell. Probes 10:159-164. - PubMed
-
- Black, S., H. Shinefield, B. Fireman, E. Lewis, P. Ray, J. R. Hansen, L. Elvin, K. M. Ensor, J. Hackell, G. Siber, F. Malinoski, D. Madore, I. Chang, R. Kohberger, W. Watson, R. Austrian, K. Edwards, et al. 2000. Efficacy, safety and immunogenicity of heptavalent pneumococcal conjugate vaccine in children. Pediatr. Infect. Dis. J. 19:187-195. - PubMed
-
- Centers for Disease Control and Prevention. 1997. Prevention of pneumococcal disease: recommendations of the Advisory Committee on Immunization Practices (ACIP). Morb. Mortal. Wkly. Rep. 46:1-24. - PubMed
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources
Molecular Biology Databases