title: A tetra-primer amplification refractory mutation system polymerase chain reaction for the evaluation of rs12979860 IL28B genotype.
abstract: Recently, genome-wide association studies in patients affected by HCV infection have identified a strong association between sustained virological response to peg-interferon/ribavirin and spontaneous viral clearance and common single nucleotide polymorphisms (SNPs) near the IL28B gene, encoding for interferon-lambda-3. Thus, it is anticipated that IL28B genotype determination will be integrated in clinical practice to guide treatment decisions. Here, we describe a simple tetra-primer amplification refractory mutation system polymerase chain reaction (T-ARMS-PCR) for the evaluation of the rs12979860 C>T IL28B SNP, for which strong evidence of association with clinical outcomes has been collected in subjects of European descent. Valid genotypic data were obtained for over 99% of subjects analysed, and T-ARMS-PCR procedures were validated by the analysis of DNA samples of 164 patients with chronic HCV infection. In conclusion, this method allows rapid, reproducible, inexpensive and accurate detection of rs12979860 polymorphism without need of any special equipment and is also suitable for evaluation of a low number of samples on a routine basis.
authors.affiliation: Department of Internal Medicine, Università degli Studi, Ospedale Maggiore Policlinico Ca' Granda IRCCS, Milano, Italy.
authors.forename: E
authors.initials: E
authors.lastname: Galmozzi
authors.forename: B
authors.initials: B
authors.lastname: Del Menico
authors.forename: R
authors.initials: R
authors.lastname: Rametta
authors.forename: P
authors.initials: P
authors.lastname: Dongiovanni
authors.forename: A L
authors.initials: AL
authors.lastname: Fracanzani
authors.forename: L
authors.initials: L
authors.lastname: Benedan
authors.forename: V
authors.initials: V
authors.lastname: Borroni
authors.forename: P
authors.initials: P
authors.lastname: Maggioni
authors.forename: S
authors.initials: S
authors.lastname: Fargion
authors.forename: L
authors.initials: L
authors.lastname: Valenti
mesh_terms: D000328:Adult; D000368:Aged; D000483:Alleles; D000998:Antiviral Agents / Q000627:therapeutic use; D005260:Female; D005820:Genetic Testing / Q000379:methods*; D005838:Genotype; D016174:Hepacivirus / Q000472:pathogenicity; D019698:Hepatitis C, Chronic / Q000188:drug therapy / Q000235:genetics* / Q000821:virology; D006801:Humans; D007372:Interferons; D007378:Interleukins / Q000235:genetics* / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D016133:Polymerase Chain Reaction / Q000379:methods*; D020641:Polymorphism, Single Nucleotide; D012254:Ribavirin / Q000627:therapeutic use; D016896:Treatment Outcome; D044465:White People / Q000235:genetics; D000095022:Interferon Lambda
chemical_list: D000998:Antiviral Agents; D007372:Interferons; D007378:Interleukins; D012254:Ribavirin; C000624353:interferon-lambda, human; D000095022:Interferon Lambda