Quadruplex Genotyping of TLR-8 Polymorphisms in a Single Reaction

Abstract

Toll-like Receptor 8 (TLR8) plays an important role in the innate immune defense against various pathogens. The TLR8 genetic variation affects the course of infections with agents such as HIV, HCV, and mycobacteria. To assess the influence of single nucleotide polymorphisms (SNPs) large cohorts need to be studied; thus, rapid and cost-efficient protocols for high-throughput TLR8 genotyping are needed. We designed a single-tube assay for genotyping four SNPs located to the major coding exon of TLR8 using the LightCycler 480 system. The new method is accurate, fast, low-priced, and can be applied for fully automated high-throughput genotyping of TLR8 SNPs.

Keywords:

• Innate immunity
• Lightcycler
• Single tube assay
• SNP
• TLR

We thank Fränzi Creutzburg and Diana Woellner for outstanding technical assistance. This work forms part of the doctoral theses of A.S. and K.B. It was supported by the Charité – Universitätsmedizin Berlin (Rahel Hirsch Grant to D.-Y. O.) and by the Deutsche Forschungsgesellschaft (grants to RRS).

The authors declare no competing interests.

Notes

¹All nucleic acid positions refer to the start codon of TLR8 mRNA with the reference number NM_138636.

²Nucleic acid positions refer to the start codon of TLR8 mRNA reference number NM_138636.

^a Derived from the pUNO or the pcDNA vector, respectively, by inducing the particular mutation (bold) to the indicated site.

³Site of mutation bold.