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Author Spotlight: Advancing Rapid Detection of Respiratory Pathogens Using Microfluidic Chip
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Molecular serotype-specific identification of Streptococcus pneumoniae using loop-mediated isothermal amplification
Chika Takano1,2, Yoko Kuramochi3, Mitsuko Seki4,5
1Division of Microbiology, Department of Pathology and Microbiology, Nihon University School of Medicine, Tokyo, 173-8610, Japan.
Scientific Reports
|December 29, 2019
Summary
A new loop-mediated isothermal amplification (LAMP) assay rapidly identifies Streptococcus pneumoniae serotypes targeted by the pneumococcal conjugate vaccine (PCV13). This cost-effective method offers improved sensitivity for detecting pneumococcal serotypes in resource-limited settings.
Area of Science:
- Microbiology
- Molecular Biology
- Vaccinology
Background:
- Pneumococcal meningitis incidence decreased with PCV7 and PCV13 introduction.
- Emergence of non-PCV13 serotypes in developed countries.
- PCV13-targeted serotypes remain a public health concern in resource-limited settings.
Purpose of the Study:
- Develop a rapid, simple, and cost-effective assay for Streptococcus pneumoniae serotyping.
- Create a novel loop-mediated isothermal amplification (LAMP) assay for PCV13 serotypes.
- Evaluate the performance of the developed LAMP assay.
Main Methods:
- Designed a LAMP assay targeting 13 PCV13 serotypes.
- Evaluated assay reactivity, specificity, and sensitivity.
- Compared LAMP with conventional PCR assays.
- Assessed detection limits using spiked cerebrospinal fluid (CSF) and blood specimens.
Main Results:
- The LAMP assay demonstrated high specificity against 41 pneumococcal serotypes.
- LAMP sensitivity was 10-100 copies/reaction, superior to PCR (10-10^4 copies/reaction).
- Detection limits in spiked CSF and blood were comparable to purified DNA.
Conclusions:
- A rapid, simple, and sensitive LAMP assay for PCV13 serotype identification was developed.
- This is the first LAMP method for PCV13 serotype-specific identification.
- The assay shows promise for facilitating epidemiological studies of pneumococcal serotyping.
