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Giornale Italiano di Dermatologia e Venereologia 2016 October;151(5):463-6


lingua: Inglese

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

Massimiliano BERGALLO 1, 2, Renata PONTI 3, Stefano GAMBARINO 1, Ilaria GALLIANO 1, 2, Paola MONTANARI 1, 2, Paolo FAVA 3, Mauro NOVELLI 3, Pietro QUAGLINO 3, Maria T. FIERRO 3, Elena MARRA 3

1 Department of Public Health and Pediactrics, Citoimmunodiagnostic Laboratory, University of Turin, Turin, Italy; 2 Department of Pediactrics Science, Città della Salute e della Scienza di Torino, Turin, Italy; 3 Department of Medical Sciences, University of Turin, Turin, Italy


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.

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