title: Novel association of FCGR2A polymorphism with age-related macular degeneration (AMD) and development of a novel CFH real-time genotyping method.
abstract: BACKGROUND
  Age-related macular degeneration (AMD) is a degenerative ocular disease, which may lead to loss of central vision. In Caucasian populations, a strong correlation has been established with polymorphism Y402H (rs1061170) in the complement factor H gene (CFH). The H131R polymorphism (rs1801274) in the FCGR2A gene has been associated with many inflammatory diseases, but has not been investigated in relation to AMD. The goal of our study was the development of a novel method for Y402H (g.43097C>T) genotyping, the confirmation of its association with AMD in the Greek population and the investigation of the H131R polymorphism in AMD.
  
  
  METHODS
  DNAs were extracted from blood samples of 120 patients with the severe wet form of AMD and 103 age- and sex-matched controls, all of whom were clinically evaluated. A real-time PCR and melting curve analysis method for Y402H genotyping was developed in the LightCycler platform, after in silico design of appropriate primers and probes. Genotyping for H131R was performed using a real-time PCR method previously described by our group.
  
  
  RESULTS
  The novel genotyping method for Y402H in the CFH gene is fast, reproducible (Efficiency=1.79, reproducibility CVCq=3.33%, Tm C allele 53.36 °C and T allele 61.91 °C, ΔTm=8.55) and accurate as results were confirmed with the gold standard DNA Sequencing method.
  
  
  CONCLUSIONS
  The present study confirmed the association between CFH Y402H SNP and wet AMD in the Greek population (OR=1.77, p=0.002). FCGR2A H131R polymorphism was investigated for the first time in this present study for possible correlation with wet AMD and a statistically significant association was detected (OR=1.74, p=0.006), that awaits further confirmation in a larger set of samples.
authors: Aliki
authors: A
authors: Velissari
authors: Iosif
authors: I
authors: Skalidakis
authors: Samantha C
authors: SC
authors: Oliveira
authors: Chryssanthi
authors: C
authors: Koutsandrea
authors: George
authors: G
authors: Kitsos
authors: Michael B
authors: MB
authors: Petersen
authors: Christos
authors: http://orcid.org/0000-0002-5876-2599
authors: C
authors: Kroupis
mesh_terms: D000368:Aged; D000483:Alleles; D016022:Case-Control Studies; D017242:Complement Factor H / Q000235:genetics*; D005260:Female; D060005:Genotyping Techniques / Q000379:methods; D006801:Humans; D008268:Macular Degeneration / Q000097:blood / Q000235:genetics*; D008875:Middle Aged; D020641:Polymorphism, Single Nucleotide; D060888:Real-Time Polymerase Chain Reaction; D017452:Receptors, IgG / Q000097:blood / Q000235:genetics*; D015203:Reproducibility of Results; D017422:Sequence Analysis, DNA; D044465:White People
chemical_list: C445323:FCGR2A protein, human; D017452:Receptors, IgG; D017242:Complement Factor H