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Measles virus genotyping by nucleotide-specific multiplex PCR
Jacques R Kremer1, Fred Fack, Christophe M Olinger
1Department of Immunology and WHO Collaborative Centre for Measles, Laboratoire National de Santé, 20A, rue Auguste Lumière, L-1011 Luxembourg, Luxembourg.
Journal of Clinical Microbiology
|July 10, 2004
Summary
A new multiplex PCR method simplifies measles virus (MV) genotyping, replacing sequencing for faster clade and genotype identification. This approach aids global measles control efforts and WHO elimination strategies.
Area of Science:
- Virology
- Molecular Biology
- Public Health
Background:
- Measles virus (MV) genotyping is crucial for tracking transmission and guiding elimination strategies.
- Current sequencing methods can be time-consuming and resource-intensive, particularly in resource-limited settings.
- Accurate and rapid genotyping supports the World Health Organization's (WHO) measles control goals.
Purpose of the Study:
- To develop a simple, rapid, and cost-effective method for measles virus (MV) genotyping.
- To replace the conventional sequencing reaction with multiplex PCR for clade and genotype identification.
- To facilitate MV genotyping for enhanced global measles control and elimination.
Main Methods:
- Developed a genotyping method using multiplex PCR, replacing sequencing.
- Utilized six multiplex PCRs: one for clade and five for genotype identification.
- Designed primers specific to MV clades and genotypes, analyzing fragment sizes via agarose gel electrophoresis.
Main Results:
- The multiplex PCR method successfully discriminated between all active MV clades and genotypes.
- Calculated positive and negative predictive values of 99.2% and 98.6%, respectively, based on published MV sequences.
- Demonstrated limited variability in primer binding sites, ensuring high sensitivity.
Conclusions:
- The developed multiplex PCR method offers a viable alternative to sequencing for MV genotyping.
- This technique is particularly beneficial for measles virus genotyping in developing countries.
- The method will contribute to improved MV control and support WHO-recommended elimination strategies.