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Simultaneous detection of Streptococcus pneumoniae and Haemophilus influenzae by nested PCR amplification from
K Saruta1, T Matsunaga, M Kono
1Department of Laboratory Medicine, Jikei University School of Medicine, Tokyo, Japan.
Abstract:
Haemophilus influenzae and Streptococcus pneumoniae are often the cause of serious diseases such as meningitis. We designed a nested PCR assay to identify these pathogens from cerebrospinal fluid samples. The first-step PCR was able to detect eubacterial rRNA genes with a unified set of universal primers. In the second-step PCR, the identification primers, HI I and II and SP I and II, could detect H. influenzae and S. pneumoniae respectively through amplification of the rRNA spacer between the 16S and 23S rRNA genes. We suggest that the two-step PCR assay can be used as a novel method for the immediate and retrospective diagnosis of bacterial meningitis caused by H. influenzae and S. pneumoniae.
Insights
A novel nested PCR assay effectively identifies Haemophilus influenzae and Streptococcus pneumoniae, common causes of meningitis. This method offers a new approach for diagnosing bacterial meningitis from cerebrospinal fluid samples.
Area of Science:
- Microbiology
- Molecular Biology
- Infectious Diseases
Background:
- Haemophilus influenzae and Streptococcus pneumoniae are significant bacterial pathogens.
- These bacteria are leading causes of serious infections, including meningitis.
- Accurate and rapid diagnostic methods are crucial for effective treatment.
Purpose of the Study:
- To develop and evaluate a nested PCR assay for identifying H. influenzae and S. pneumoniae.
- To assess the utility of the assay in cerebrospinal fluid (CSF) samples.
- To provide a novel diagnostic tool for bacterial meningitis.
Main Methods:
- A two-step nested PCR assay was designed.
- The first step used universal primers to detect eubacterial rRNA genes.
- The second step employed specific primers (HI I/II and SP I/II) to identify H. influenzae and S. pneumoniae by amplifying the rRNA spacer region.
Main Results:
- The nested PCR assay successfully detected H. influenzae and S. pneumoniae.
- The assay demonstrated capability in identifying these pathogens from CSF samples.
- The method targets the rRNA spacer region for specific pathogen identification.
Conclusions:
- The developed two-step PCR assay is a viable method for bacterial meningitis diagnosis.
- This assay can be used for both immediate and retrospective diagnosis.
- It offers a novel approach for identifying H. influenzae and S. pneumoniae in meningitis cases.