A < i > Borrelia burgdorferi < /i > outer surface protein C (OspC) genotyping method using Luminex technology

by Patrick Pearson, Olivia Skaltsis, Chu-Yuan Luo, Guang Xu, Zachary Oppler, Dustin Brisson, Stephen M. RichBorrelia burgdorferi is an important tickborne human pathogen comprising several strains based on nucleotide sequence of the outer surface protein C (ospC) gene. Detection and characterization of differentospC genotypes is vital for research onB.burgdorferi and the risk it poses to humans. Here we present a novel, multiplex assay based on Luminex xMAP technology for the detection ofB.burgdorferi ospC genotypes. The assay has five major steps: amplification of theospC gene, hydrolyzation of surplus primers and nucleotides, incorporation of biotinylated nucleotides into the template DNA, hybridization to Luminex microspheres, and detection of fluorescent signals corresponding to eachospC genotype. We validated the protocol by comparing results obtained from our method against results from an establishedospC genotyping method. This protocol can be used for the characterization ofospC genotypes inB.burgdorferi infected ticks, reservoir hosts, and/or clinical samples.
Source: PLoS One - Category: Biomedical Science Authors: Source Type: research