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Published on: October 24, 2010
Molecular methods for the detection and characterization of Neisseria meningitidis
Mathew A Diggle1, Stuart C Clarke
1Stobhill Hospital, Scottish Meningococcus & Pneumococcus Reference Laboratory, Glasgow, UK.
Abstract:
Neisseria meningitidis remains a common global cause of morbidity and mortality. The laboratory confirmation of meningococcal disease is, therefore, very important for individual patient management and for public health management. Through surveillance schemes, it provides long-term epidemiologic data that can be used to inform vaccine policy. Traditional methods, such as latex agglutination and the enzyme-linked immunosorbent assay, are still used, but molecular methods are now also established. In this review, molecular methods for the laboratory confirmation and characterization of meningococci are described. PCR is an invaluable tool in modern biology and can be used to predict the group, type and subtype of meningococci. It is now also used in a fluorescence-based format for increased sensitivity and specificity. The method also provides the amplified DNA for other techniques, such as multilocus sequence typing. Other methods for the discrimination of meningococci have also played and continue to play an important part in epidemiology. For example, pulsed-field gel electrophoresis is highly discriminatory, whilst multilocus enzyme electrophoresis provided the basis for the description of global meningococcal clones and formed the foundation for multilocus sequence typing. Other less commonly used methods, such as matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and pyrosequencing, may increasingly find their way into microbiology reference laboratories. Nevertheless, nucleotide sequencing and laboratory automation have aided the introduction of many methods and provide data that are digitally based and, therefore, highly accurate and portable.
Insights
Laboratory confirmation of meningococcal disease is crucial for patient and public health. Molecular methods like PCR offer accurate identification and characterization of Neisseria meningitidis, aiding surveillance and vaccine policy.
Area of Science:
- Microbiology
- Epidemiology
- Molecular Biology
Background:
- Neisseria meningitidis is a significant cause of global morbidity and mortality.
- Accurate laboratory confirmation is vital for disease management, public health surveillance, and informing vaccine policy.
- Traditional diagnostic methods are complemented by established molecular techniques.
Purpose of the Study:
- To review molecular methods for laboratory confirmation and characterization of Neisseria meningitidis.
- To highlight the utility of various molecular techniques in epidemiological studies and public health.
Main Methods:
- Polymerase Chain Reaction (PCR) for group, type, and subtype prediction.
- Fluorescence-based PCR for enhanced sensitivity and specificity.
- Multilocus Sequence Typing (MLST) utilizing amplified DNA from PCR.
- Pulsed-Field Gel Electrophoresis (PFGE) for high discriminatory power.
- Multilocus Enzyme Electrophoresis (MLEE) for defining meningococcal clones.
- Emerging techniques: MALDI-TOF MS and pyrosequencing.
Main Results:
- PCR is a versatile tool for meningococcal identification and characterization.
- Molecular methods provide accurate, portable, and digitally based epidemiological data.
- Advanced techniques like PFGE and MLEE have been foundational in understanding meningococcal epidemiology and clonal structures.
Conclusions:
- Molecular methods are essential for accurate laboratory confirmation and characterization of Neisseria meningitidis.
- These techniques significantly contribute to disease surveillance, public health management, and vaccine policy development.
- Nucleotide sequencing and laboratory automation are key drivers in the adoption and accuracy of modern microbiological methods.
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