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A quantitative LightCycler PCR to detect Streptococcus pneumoniae in blood and CSF
Robert van Haeften1, Silvano Palladino, Ian Kay
1Department of Microbiology and Infectious Diseases, Royal Perth Hospital, Perth, Australia. robert.vanhaeften@health.wa.gov.au
Diagnostic Microbiology and Infectious Disease
|October 3, 2003
Summary
A new quantitative PCR test accurately detects Streptococcus pneumoniae using Pneumolysin gene detection. This sensitive assay shows potential for rapid diagnosis and improved patient management of pneumococcal infections.
Area of Science:
- Molecular Biology
- Clinical Microbiology
- Infectious Diseases
Background:
- Streptococcus pneumoniae is a leading cause of invasive bacterial infections.
- Rapid and accurate diagnostic methods are crucial for timely treatment.
- Current diagnostic methods can be slow and lack sensitivity.
Purpose of the Study:
- To develop and validate a quantitative real-time PCR assay for detecting Streptococcus pneumoniae.
- To evaluate the assay's sensitivity, specificity, and performance in clinical specimens.
- To assess the potential of the assay for improving patient management.
Main Methods:
- Development of a quantitative real-time PCR assay targeting the Pneumolysin (ply) gene.
- Utilized Fluorescence Resonance Energy Transfer (FRET) probes on the LightCycler instrument.
- Tested the assay on cerebrospinal fluid (CSF) and whole blood specimens from patients with invasive pneumococcal disease.
Main Results:
- The assay detected all common S. pneumoniae serotypes with high sensitivity (1-10 target copies/reaction).
- 100% of CSF and 69% of whole blood specimens from patients were PCR positive.
- High specificity was observed, with negative results in non-pneumococcal samples and healthy controls.
Conclusions:
- The developed PCR assay is highly sensitive and specific for Streptococcus pneumoniae detection.
- The assay provides rapid results, potentially improving antibiotic treatment for pneumococcal infections.
- Bacterial load quantitation offers new possibilities for patient management and monitoring.