title: A novel TaqMAMA assay for allelic discrimination of TLR9 rs352140 polymorphism.
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.
authors.affiliation: Department of Public Health and Pediatric Sciences, University of Turin, Medical School, 10136, Turin, Italy. Electronic address: massimiliano.bergallo@unito.it.
authors.forename: Massimiliano
authors.initials: M
authors.lastname: Bergallo
authors.affiliation: Department of Public Health and Pediatric Sciences, University of Turin, Medical School, 10136, Turin, Italy.
authors.forename: Paola
authors.initials: P
authors.lastname: Montanari
authors.affiliation: Pediatric Oncohematology, Stem Cell Transplantation and Cell Therapy Division, A.O.U. Città della Salute e della Scienza-Regina Margherita Children Hospital, Turin, Italy.
authors.forename: Katia
authors.initials: K
authors.lastname: Mareschi
authors.affiliation: Department of Public Health and Pediatric Sciences, University of Turin, Medical School, 10136, Turin, Italy.
authors.forename: Marco
authors.initials: M
authors.lastname: Rassu
authors.affiliation: Department of Public Health and Pediatric Sciences, University of Turin, Medical School, 10136, Turin, Italy.
authors.forename: Ilaria
authors.initials: I
authors.lastname: Galliano
authors.affiliation: Laboratory of Molecular Virology, Azienda Ospedaliero-Universitaria Maggiore della Carità, Novara, Italy.
authors.forename: Paolo
authors.initials: P
authors.lastname: Ravanini
mesh_terms: D000328:Adult; D000368:Aged; D000483:Alleles; D003586:Cytomegalovirus Infections / Q000235:genetics* / Q000276:immunology; D005260:Female; D056726:Genetic Association Studies; D020022:Genetic Predisposition to Disease; D060005:Genotyping Techniques / Q000379:methods*; D006801:Humans; D016030:Kidney Transplantation; D008297:Male; D008875:Middle Aged; D020641:Polymorphism, Single Nucleotide; D051217:Toll-Like Receptor 9 / Q000235:genetics*; D066027:Transplant Recipients
chemical_list: C495371:TLR9 protein, human; D051217:Toll-Like Receptor 9
keywords: Allelic discrimination; PCR real time; SNPs; TLR9; TaqMAMA