Multiplexed genotyping of Bacillus anthracis by Luminex xMap suspension array.
The Luminex® xTAG technology is a medium to high throughput, open methodology able to test many single nucleotide polymorphisms (SNPs) in a single reaction and a minimum time. Multiplex SNPs interrogation are conducted on the Luminex xMAP system, which uses lasers to read universal tag, color-coded microspheres that attach to specific nucleic acid sequences. The present method describes a Multiplex Oligonucleotide Ligation-PCR procedure (MOL-PCR) for the simultaneous interrogation of 13 phylogenetically informative SNPs within the genome of Bacillus anthracis. The reported 13-plex assay enables efficient B. anthracis genotyping into major sublineages and groups. While cost-effective compared to other monoplex methods, the present MOL-PCR method also offers a high degree of flexibility and scalability. It can easily accommodate newly identified SNPs to increase resolving power to the canSNP typing of B. anthracis.
Anses, Animal Health Laboratory, Bacterial Zoonosis Unit, University Paris-Est, 23 avenue du Général De Gaulle, 94706, Maisons-Alfort, France, simon.thierry@anses.fr.
Simon
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Thierry
Sylviane
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Derzelle
D000818:Animals; D000881:Anthrax / Q000175:diagnosis / Q000382:microbiology; D001408:Bacillus anthracis / Q000145:classification / Q000235:genetics*; D006801:Humans; D058889:Molecular Typing / Q000379:methods*; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D020641:Polymorphism, Single Nucleotide