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Updated: Aug 7, 2026
Multiplex PCR Assay for Typing of Staphylococcal Cassette Chromosome Mec Types I to V in Methicillin-resistant Staphylococcus aureus
Published on: September 5, 2013
Rapid identification of Streptococcus pneumoniae serotypes by cpsB gene-based sequetyping combined with multiplex PCR
Meng-Lan Zhou1, Zi-Ran Wang1, Yan-Bing Li1
1Department of Clinical Laboratory, State Key Laboratory of Complex Severe and Rare Diseases, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China; Beijing Key Laboratory for Mechanisms Research and Precision Diagnosis of Invasive Fungal Diseases, Beijing, China.
Background/Purpose:
Streptococcus pneumoniae is an important human pathogen that causes invasive infections in adults and children. Accurate serotyping is important to study its epidemiological distribution and to assess vaccine efficacy.
Methods:
Invasive S. pneumoniae isolates (n = 300) from 27 teaching hospitals in China were studied. The Quellung reaction was used as the gold standard to identify the S. pneumoniae serotypes. Subsequently, multiplex PCR and cpsB gene-based sequetyping methods were used to identify the serotypes.
Results:
Based on the Quellung reaction, 299 S. pneumoniae isolates were accurately identified to the serotype level and 40 different serotypes were detected. Only one strain was non-typeable, and five most common serotypes were identified: 23F (43, 14.3%), 19A (41, 13.7%), 19F (41, 13.7%), 3 (31, 10.3%), and 14 (27, 9.0%). Overall, the multiplex PCR method identified 73.3 and 20.7% of the isolates to the serotype and cluster levels, respectively, with 1.7% of the isolates misidentified. In contrast, the cpsB sequetyping method identified 59.0 and 30.3% of the isolates to the serotype and cluster levels, respectively, and 7% were misidentified.
Conclusions:
The cpsB gene sequetyping method combined with multiplex PCR, can greatly improve the accuracy and efficiency of serotyping, besides reducing the associated costs.
Insights
Accurate Streptococcus pneumoniae serotyping is crucial for understanding disease spread and vaccine effectiveness. Combining multiplex PCR and cpsB gene sequencing offers a more accurate and cost-effective method for identifying pneumococcal serotypes.
Area of Science:
- Microbiology
- Infectious Diseases
- Molecular Diagnostics
Background:
- Streptococcus pneumoniae is a major cause of invasive infections in both children and adults.
- Accurate serotyping of S. pneumoniae is essential for epidemiological surveillance and evaluating vaccine impact.
Purpose of the Study:
- To compare the accuracy and efficiency of multiplex PCR and cpsB gene sequencing for S. pneumoniae serotyping against the gold standard Quellung reaction.
- To identify the most prevalent S. pneumoniae serotypes in invasive isolates from China.
Main Methods:
- Invasive S. pneumoniae isolates (n=300) from 27 Chinese hospitals were analyzed.
- Serotyping was performed using the gold standard Quellung reaction, multiplex PCR, and cpsB gene sequencing.
Main Results:
- The Quellung reaction identified 40 distinct serotypes among 299 isolates, with serotypes 23F, 19A, 19F, 3, and 14 being the most common.
- Multiplex PCR achieved serotype and cluster identification for 73.3% and 20.7% of isolates, respectively, with 1.7% misidentification.
- CpsB sequencing identified serotypes and clusters for 59.0% and 30.3% of isolates, respectively, with 7% misidentification.
Conclusions:
- Multiplex PCR and cpsB gene sequencing show promise for S. pneumoniae serotyping.
- Combining multiplex PCR with cpsB gene sequencing enhances accuracy and efficiency while reducing costs for serotyping.
- These improved methods can aid in the epidemiological study of Streptococcus pneumoniae infections.
