title: Evaluation of IFN-γ polymorphism+874 T/A in patients with recurrent tonsillitis by PCR real time mismatch amplification mutation assay (MAMA real time PCR).
abstract: Interferon gamma (IFN-γ) is an important cytokine that plays a crucial role in the balance between normal and pathological immune response. Defect of IFN-γ can give a predisposition to infectious disease, autoimmune pathologies and tumours. Different polymorphisms in this gene have been described, in particular the single nucleotide polymorphism (SNP)+874∗T/A that may affect IFN-γ gene expression. Several techniques can be used for the detection of SNPs. In this work two PCR Real Time assays were developed, an Amplification Refractory Mutation System (ARMS) and a Mismatch Amplification Mutation Assay (MAMA). Twenty-seven samples from patients (tonsillectomy) and 85 from donor's blood bank were considered. As a result, 78/85 controls (91.7%) and 25/27 patients (92.6%) were heterozygosis, considering the ARMS-PCR; 55/85 (64.7%) and 14/27 (51.9%) were heterozygosis using MAMA-PCR assay. Fourteen of 85 (16.5%) and 8/27 (29.6%) were homozygosis A, 16/85 (18.8%) and 5/27 (18.5%) presented homozygosis T, taking into account the MAMA-PCR. There are statistically difference between the two assay with p<0.0001 at Chi-square test. Our preliminary data suggest that tonsillectomy patients had a statistical trend to possess the low IFN-γ polymorphism when compared with control subject (p=0.3) but is not statistically significant. In conclusion the Real time MAMA-PCR assay has several advantages over other SNP identification techniques such as rapidity, reliability, easily to perform in one working day and applicable in clinical molecular diagnostic laboratories, although sequencing remains the gold standard.
authors.affiliation: Department of Public Health and Pediatrics, University of Turin, Italy. Electronic address: massimiliano.bergallo@unito.it.
authors.forename: Massimiliano
authors.initials: M
authors.lastname: Bergallo
authors.affiliation: Serology Unit OIRM S. Anna Hospital of Turin, Turin, Italy. Electronic address: stefano.gambarino@unito.it.
authors.forename: Stefano
authors.initials: S
authors.lastname: Gambarino
authors.affiliation: Department of Public Health and Pediatrics, University of Turin, Italy. Electronic address: elisa.loiacono@libero.it.
authors.forename: Elisa
authors.initials: E
authors.lastname: Loiacono
authors.affiliation: Department of Public Health and Pediatrics, University of Turin, Italy. Electronic address: lucavergano1987@gmail.com.
authors.forename: Luca
authors.initials: L
authors.lastname: Vergano
authors.affiliation: Department of Public Health and Pediatrics, University of Turin, Italy. Electronic address: ilaria.galliano@unito.it.
authors.forename: Ilaria
authors.initials: I
authors.lastname: Galliano
authors.affiliation: Department of Public Health and Pediatrics, University of Turin, Italy. Electronic address: paola.montanari@unito.it.
authors.forename: Paola
authors.initials: P
authors.lastname: Montanari
authors.affiliation: Istituto Zooprofilattico Sperimentale del Piemonte, Liguria e Valle D'Aosta, Turin, Italy. Electronic address: sara.astegiano@unito.it.
authors.forename: Sara
authors.initials: S
authors.lastname: Astegiano
authors.affiliation: Unit of Otorhinolaryngology, Pediatric Hospital "Regina Margherita S. Anna", Torino, Italy. Electronic address: ptavormina@cittadellasalute.to.it.
authors.forename: Paolo
authors.initials: P
authors.lastname: Tavormina
authors.affiliation: Department of Public Health and Pediatrics, University of Turin, Italy. Electronic address: pierangelo.tovo@unito.it.
authors.forename: Pier-Angelo
authors.initials: PA
authors.lastname: Tovo
mesh_terms: D000483:Alleles; D016022:Case-Control Studies; D016207:Cytokines / Q000378:metabolism; D004252:DNA Mutational Analysis / Q000379:methods*; D020022:Genetic Predisposition to Disease; D005838:Genotype; D006579:Heterozygote; D006720:Homozygote; D006801:Humans; D007249:Inflammation / Q000473:pathology; D007371:Interferon-gamma / Q000235:genetics*; D017354:Point Mutation; D011110:Polymorphism, Genetic; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012008:Recurrence; D014069:Tonsillitis / Q000235:genetics*
chemical_list: D016207:Cytokines; D007371:Interferon-gamma
keywords: ARMS-PCR; Gene expression; IFN-γ polymorphisms; MAMA-PCR; Tonsillectomy