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Multiplexed genotyping of Bacillus anthracis by Luminex xMap suspension array
Simon Thierry1, Sylviane Derzelle
1Anses, Animal Health Laboratory, Bacterial Zoonosis Unit, University Paris-Est, 23 avenue du Général De Gaulle, 94706, Maisons-Alfort, France, simon.thierry@anses.fr.
Methods in Molecular Biology (Clifton, N.J.)
|November 17, 2014
Summary
This study introduces a cost-effective Multiplex Oligonucleotide Ligation-PCR (MOL-PCR) method for rapid Bacillus anthracis genotyping. The assay efficiently identifies major B. anthracis sublineages using 13 single nucleotide polymorphisms (SNPs).
Area of Science:
- Molecular Biology
- Genetics
- Microbiology
Background:
- Single nucleotide polymorphisms (SNPs) are crucial for bacterial genotyping.
- High-throughput methods are needed for efficient analysis of multiple SNPs.
- Bacillus anthracis genotyping is important for tracking outbreaks and understanding its evolution.
Purpose of the Study:
- To develop and validate a Multiplex Oligonucleotide Ligation-PCR (MOL-PCR) assay.
- To simultaneously interrogate 13 phylogenetically informative SNPs in Bacillus anthracis.
- To enable efficient and scalable genotyping of B. anthracis.
Main Methods:
- Utilized Luminex® xTAG technology and the Luminex xMAP system.
- Developed a 13-plex Multiplex Oligonucleotide Ligation-PCR (MOL-PCR) procedure.
- Applied the assay to interrogate SNPs within the Bacillus anthracis genome.
Main Results:
- The MOL-PCR assay successfully genotyped Bacillus anthracis.
- Efficient differentiation into major sublineages and groups was achieved.
- The method demonstrated cost-effectiveness, flexibility, and scalability.
Conclusions:
- The developed 13-plex MOL-PCR assay provides an efficient tool for Bacillus anthracis genotyping.
- This method offers a cost-effective and scalable alternative to monoplex approaches.
- The assay can be adapted to include new SNPs for enhanced typing resolution.
