Accurate serotype identification of Streptococcus pneumoniae using nanopore Cas9-targeted serotype identification (nCATSerotyping)
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View abstract on PubMed
Summary
This summary is machine-generated.A new nanopore Cas9-targeted serotyping (nCATSerotyping) platform accurately identifies Streptococcus pneumoniae serotypes, including emerging ones. This method significantly outperforms conventional techniques for epidemiological surveillance and vaccine development.
Area Of Science
- Microbiology and Infectious Diseases
- Genomics and Bioinformatics
- Vaccinology
Background
- Streptococcus pneumoniae causes significant community-acquired pneumonia and invasive diseases globally.
- Pneumococcal conjugate vaccines have reduced disease but led to the rise of non-vaccine and emerging serotypes.
- Conventional serotyping methods (mPCR, mAb, Quellung) have limitations in coverage and accuracy for novel serotypes.
Purpose Of The Study
- To develop and validate a novel nanopore Cas9-targeted serotyping (nCATSerotyping) platform for accurate S. pneumoniae identification.
- To assess the platform's performance against conventional methods using clinical isolates.
- To enable improved epidemiological surveillance and next-generation vaccine development.
Main Methods
- Development of the nCATSerotyping platform utilizing Cas9-mediated enrichment and Oxford Nanopore sequencing.
- Application of nCATSerotyping to 276 clinical S. pneumoniae isolates from South Korea (2018-2020).
- Comparative analysis with monoclonal antibody (mAb) and multiplex PCR (mPCR) methods; whole-genome sequencing for non-typeable isolates.
Main Results
- nCATSerotyping achieved a 97.10% success rate (268/276 isolates), significantly higher than mAb and mPCR (76.45%, 211/276).
- Whole-genome sequencing confirmed 100% accuracy for S. pneumoniae serotyping, identifying the remaining isolates as oral streptococci.
- The platform successfully identified emerging serotypes (e.g., serotype 13) and null capsule clade strains missed by conventional methods.
Conclusions
- nCATSerotyping provides a rapid, accurate, and comprehensive solution for S. pneumoniae serotyping, excelling with novel and non-typeable strains.
- The platform demonstrates significant advantages over conventional methods, crucial for accurate epidemiological surveillance.
- nCATSerotyping is a scalable tool essential for monitoring vaccine effectiveness and guiding next-generation multivalent pneumococcal vaccine development.
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