Accurate, fast and cost-effective simultaneous detection of bacterial meningitis by qualitative PCR with high-resolution melting.
Ivano de FilippisAline Carvalho de AzevedoIrene de Oliveira LimaNicolle F Lima RamosClaudia Ferreira de AndradeAntonio Ecc de AlmeidaPublished in: BioTechniques (2023)
Neisseria meningitidis , Haemophilus influenzae and Streptococcus pneumoniae are leading causes of meningitis and acute invasive infections. PCR-based methods are widely used for the diagnosis and surveillance of bacterial pathogens because of their high sensitivity, specificity and high-throughput capabilities compared with conventional laboratory methods. This study evaluated a high-resolution melting qualitative PCR analysis method for the simultaneous detection of these three pathogens. The assay has been optimized to detect three species-specific genes of each organism isolated from clinical samples, enabling accurate identification of the etiological agent. The method proved to be highly sensitive and cheaper than the real-time PCR TaqMan ® system because it is probe-free; it could be used for the diagnosis of invasive diseases in public health laboratories of developing countries.
Keyphrases
- real time pcr
- high resolution
- public health
- high throughput
- mass spectrometry
- gram negative
- cerebrospinal fluid
- bioinformatics analysis
- liver failure
- antimicrobial resistance
- systematic review
- high speed
- tandem mass spectrometry
- genome wide
- gene expression
- respiratory failure
- intensive care unit
- quantum dots
- fluorescent probe
- aortic dissection
- liquid chromatography
- simultaneous determination