title: PCR Real time Mismatch Amplification Mutation Assay (MAMA Real Time PCR) for evaluation of TNF-α promoter gene polymorphism -308 G/A in patients with psoriasis.
abstract: BACKGROUND
  Psoriasis is a common chronic inflammatory disease, the plaques are infiltrated by leukocytes producing high levels of proinflammatory cytokines and TNF-α. Single-nucleotide polymorphisms within the gene promoters have been shown to affect gene expression. The -308 G/A polymorphism could affect TNF synthesis at transcriptional level. The present study develops a MAMA Real Time PCR assay, in order to identify homozygosis or heterozygosis for TNF-α -308 G/A polymorphism.
  
  
  METHODS
  Seventy patients with psoriasis and 235 controls were considered for the development of the real time PCR assay. Whole blood was processed for nucleic acid extraction.
  
  
  RESULTS
  A percentage of 36.17% controls and 38.6% patients were heterozygosis, considering Amplification-refractory mutation system (ARMS)-PCR assay while 23% and 22.85% were heterozygosis using Mismatch Amplification Mutation Assay (MAMA)-PCR. On the contrary, 1.3% and 1.4% were homozygosis A, while 75.7% and 75.75% presented homozygosis G, taking into account the MAMA-PCR results. The two assays were significantly different (P=0.0004 at χ2 Test), but MAMA-PCR showed a better performance for TNF-α -308 G/A gene polymorphism investigation.
  
  
  CONCLUSIONS
  Further studies are needed for a better comprehension of the role of this polymorphism, such as MAMA real time PCR assays development for other players in cellular immune response.
authors.affiliation: Department of Public Health and Pediactrics, Citoimmunodiagnostic Laboratory, University of Turin, Turin, Italy - massimiliano.bergallo@unito.it.
authors.forename: Massimiliano
authors.initials: M
authors.lastname: Bergallo
authors.forename: Renata
authors.initials: R
authors.lastname: Ponti
authors.forename: Stefano
authors.initials: S
authors.lastname: Gambarino
authors.forename: Ilaria
authors.initials: I
authors.lastname: Galliano
authors.forename: Paola
authors.initials: P
authors.lastname: Montanari
authors.forename: Paolo
authors.initials: P
authors.lastname: Fava
authors.forename: Mauro
authors.initials: M
authors.lastname: Novelli
authors.forename: Pietro
authors.initials: P
authors.lastname: Quaglino
authors.forename: Maria T
authors.initials: MT
authors.lastname: Fierro
authors.forename: Elena
authors.initials: E
authors.lastname: Marra
mesh_terms: D016022:Case-Control Studies; D004252:DNA Mutational Analysis / Q000379:methods; D005786:Gene Expression Regulation; D020022:Genetic Predisposition to Disease; D006801:Humans; D009154:Mutation; D020641:Polymorphism, Single Nucleotide; D011401:Promoter Regions, Genetic; D011565:Psoriasis / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics*
chemical_list: D014409:Tumor Necrosis Factor-alpha