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[Diagnostics of invasive meningococcal, haemophilus and pneumococcal disease by PCR assay]
Jitka Kalmusová1, Eva Bronská, Pavla Krízová
1National Reference Laboratory for Meningococcal Infections, National Institute of Public Health, Prague, Czech Republic, jkalm@szu.cz
Objectives:
Development of extended polymerase chain reaction (PCR) for non-culture detection of Nesseria meningitidis, Haemophilus influenzae and Streptococcus pneumonie from invasive infections.
Materials And Methods:
A method of PCR was optimalised on strains of Nesseria meningitidis, Haemophilus influenzae b and Streptococcus pneumonie. Detection of pathogens was evaluated on 230 samples from patiens with invasive infection.
Results:
Positive results of PCR were found in 103 samples of 230 (44.7 %). The percentage of positivity was higher in CSF samples (57.0 %) than in serum (33.8 %) or blood (33.3 %) samples.
Conclusion:
PCR method enables etiological diagnostics in cases, where antibiotic treatment was started. PCR results are available earlier than the results of cultivation. Multilocus sequence typing (MLST) of PCR products enables clonal analysis of etiological agents even in cases with negative results of cultivation.
Insights
This study developed an extended polymerase chain reaction (PCR) for detecting Neisseria meningitidis, Haemophilus influenzae, and Streptococcus pneumoniae in invasive infections. The PCR method offers faster etiological diagnostics than traditional cultivation, even after antibiotic treatment has begun.
Area of Science:
- Microbiology
- Molecular Biology
- Infectious Diseases
Background:
- Invasive bacterial infections caused by Neisseria meningitidis, Haemophilus influenzae, and Streptococcus pneumoniae pose significant public health challenges.
- Traditional culture-based methods for diagnosing these infections can be slow and may yield false negatives, especially after antibiotic administration.
Purpose of the Study:
- To develop and evaluate an extended polymerase chain reaction (PCR) assay for the rapid, non-culture detection of Neisseria meningitidis, Haemophilus influenzae, and Streptococcus pneumoniae.
- To assess the utility of PCR in diagnosing invasive infections, particularly in patients who have already commenced antibiotic therapy.
Main Methods:
- An extended polymerase chain reaction (PCR) method was optimized using reference strains of Neisseria meningitidis, Haemophilus influenzae type b, and Streptococcus pneumoniae.
- The optimized PCR assay was applied to 230 clinical samples (cerebrospinal fluid, serum, and blood) obtained from patients with suspected invasive infections.
Main Results:
- The PCR assay detected target pathogens in 103 out of 230 samples (44.7%).
- Detection rates varied by sample type, with higher positivity observed in cerebrospinal fluid (57.0%) compared to serum (33.8%) and blood (33.3%).
Conclusions:
- The developed PCR method provides a valuable tool for etiological diagnosis of invasive bacterial infections, especially when antibiotic treatment has been initiated.
- PCR offers significantly faster results than conventional cultivation, enabling timely clinical decisions.
- Subsequent multilocus sequence typing (MLST) of PCR products allows for clonal analysis of pathogens, even in culture-negative cases.
