A real-time PCR assay for the simultaneous detection of functional N284I and L412F polymorphisms in the human toll-like receptor 3 gene

Toll-like receptors (TLR) serve important roles in immunity against microbial pathogens, and TLR3 plays a critical role in the recognition of double-stranded RNA derived from viruses. Two specific single nucleotide polymorphisms (SNP) in the human TLR3 gene, A851T and C1234T, which result in amino acid substitutions, N284I and L412F, respectively, have been described to result in impaired receptor signaling and function in an in vitro model. Hence, an assay that could simultaneously detect these two functional TLR3 SNPs could facilitate studies into their role in the pathogenesis of human viral diseases and serve as a tool to screen viral disease predisposition. Therefore, we developed a real-time PCR assay based on a Light-Cycler format for simultaneous detection of A851T and C1234T SNPs within the TLR3 gene. This relatively low-cost PCR assay is proposed as a novel tool in the study of TLR3 and its role in viral disease.