A novel TaqMAMA assay for allelic discrimination of TLR9 rs352140 polymorphism

J Virol Methods. 2017 May:243:25-30. doi: 10.1016/j.jviromet.2017.01.015. Epub 2017 Jan 28.

Abstract

TaqMAMA is an allele-specific PCR-based (ASPCR) method that may be suitable for broad and cost-effective genotyping applications in all types of laboratories. There is evidence that interactions between some toll like receptors (TLRs) with viruses influence both the immune response and outcome of HCMV infection. We developed a TaqMAMA genotyping assay for the detection of rs352140 TLR9 polymorphism in transplant recipients with and without HCMV infections. Performance parameters to ensure a solid pre-validation protocol have been here argued. We analysed a population of 74 kidney transplants recipients subdivided in 58 HCMV PCR positive and 16 HCMV PCR negative in the post-transplant routine control. All 74 samples were tested with 31/74 (41.9%) homozygotes (11 CC and 20 TT) and 43/74 (58.1%) heterozygotes (CT). Our preliminary data suggest that there is no correlation between TLR9 rs352140 polymorphism and frequency of HCMV infection.

Keywords: Allelic discrimination; PCR real time; SNPs; TLR9; TaqMAMA.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Aged
  • Alleles*
  • Cytomegalovirus Infections / genetics*
  • Cytomegalovirus Infections / immunology
  • Female
  • Genetic Association Studies
  • Genetic Predisposition to Disease*
  • Genotyping Techniques / methods*
  • Humans
  • Kidney Transplantation
  • Male
  • Middle Aged
  • Polymorphism, Single Nucleotide*
  • Toll-Like Receptor 9 / genetics*
  • Transplant Recipients

Substances

  • TLR9 protein, human
  • Toll-Like Receptor 9