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A novel LAMP-based assay for the identification of Streptococcus pneumoniae and Streptococcus pseudopneumoniae in
Mariam Basheer1,2, Katarina Sasic2, Alice Carlsson2
1Division of Clinical Microbiology, Karolinska University Hospital, Stockholm, Sweden.
Abstract:
The Gram-positive bacteria Streptococcus pneumoniae is part of the Streptococcus mitis group (SMG) and causes life-threatening infections, such as pneumonia, sepsis, and meningitis. The closely related Streptococcus pseudopneumoniae has recently been shown to cause respiratory tract infections, as well as invasive infections, especially in patients with comorbidities. Due to the genetic and phenotypic similarities of species belonging to the SMG, the identification of S. pneumoniae and S. pseudopneumoniae is difficult and unreliable using phenotypic tests, as well as matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) and 16S rRNA sequencing. In this study, a loop-mediated isothermal amplification (LAMP)-based assay was developed using molecular markers specific for S. pneumoniae (SPN0001) and S. pseudopneumoniae (SPPN_RS10375). The LAMP assay was evaluated using a collection of SMG clinical isolates, concluding that the method provides a correct, reliable, and fast identification of both S. pneumoniae and S. pseudopneumoniae clinical isolates.
Insights
Accurate identification of Streptococcus pneumoniae and Streptococcus pseudopneumoniae is challenging. A new loop-mediated isothermal amplification (LAMP) assay offers a reliable and rapid method for distinguishing these clinically significant bacteria.
Area of Science:
- Microbiology
- Molecular Diagnostics
- Bacteriology
Background:
- Streptococcus pneumoniae and Streptococcus pseudopneumoniae are Gram-positive bacteria within the Streptococcus mitis group (SMG).
- Both species cause serious infections, including pneumonia, sepsis, and meningitis, with S. pseudopneumoniae increasingly recognized in respiratory and invasive infections, particularly in immunocompromised individuals.
- Distinguishing between S. pneumoniae and S. pseudopneumoniae is difficult due to genetic and phenotypic similarities, making traditional identification methods unreliable.
Purpose of the Study:
- To develop a novel molecular diagnostic assay for the accurate differentiation of S. pneumoniae and S. pseudopneumoniae.
- To address the limitations of current phenotypic and molecular identification techniques for SMG species.
Main Methods:
- Development of a loop-mediated isothermal amplification (LAMP)-based assay targeting specific molecular markers for S. pneumoniae (SPN0001) and S. pseudopneumoniae (SPPN_RS10375).
- Evaluation of the LAMP assay's performance using a diverse collection of clinical isolates from the SMG.
Main Results:
- The developed LAMP assay demonstrated high specificity and sensitivity for identifying both S. pneumoniae and S. pseudopneumoniae.
- The assay provided correct and reliable differentiation between the two closely related species.
Conclusions:
- The molecular LAMP assay is a fast, accurate, and reliable tool for the identification of S. pneumoniae and S. pseudopneumoniae clinical isolates.
- This method can significantly improve the diagnosis and management of infections caused by these important pathogens.
