title: Comparison of the results of PCR-RFLP and reverse hybridization methods used in molecular diagnosis of FMF.
abstract: Familial Mediterranean fever (FMF) is characterized by recurrent fever, serositis, and arthritis. Due to the abundance of mutations and clinical heterogeneity of the disease, different screening methods have been developed. In this study, we aimed to compare our findings of mutations determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) with reverse hybridization (RH) methods. In 152 of 263 patients (57.79%) different mutations were determined with RH. Allelic frequencies were E148Q 6.84%, M680I(G/C) 3.61%, M694V 20.91%, V726A 7.03%, P369S 1.33%, F479L 0.19%, M680I(G/A) 0.76%, M694I 0.57%, K695R 0.57%, A744S 0.38%, R731H 0.38%, and del1692 0%. Frequent mutations were also confirmed by PCR-RFLP. There were no conflicting results between the two methods. Four of these genotypes were homozygous for a single mutation, 15 were heterozygous for two mutations, 8 were heterozygous for a single mutation, 1 was heterozygous for three mutations, and 1 was homozygous for one mutation and heterozygous for another mutation. It has been reported that analytical sensitivity of RH is 97%. We did not find a discrepancy between the two methods. In 21 patients, we detected additional mutations with RH. This finding was regarded as an advantage of RH, and we concluded that this assay is a useful method for detection of first stage FMF mutation screening.
authors.affiliation: Department of Medical Genetics, Baskent University Faculty of Medicine, Ankara, Turkey. feridesahin@hotmail.com
authors.forename: Feride Iffet
authors.initials: FI
authors.lastname: Sahin
authors.forename: Zerrin
authors.initials: Z
authors.lastname: Yilmaz
authors.forename: Erkan
authors.initials: E
authors.lastname: Yurtcu
authors.forename: Esra
authors.initials: E
authors.lastname: Baskin
mesh_terms: D000483:Alleles; D001483:Base Sequence; D003598:Cytoskeletal Proteins / Q000235:genetics*; D017931:DNA Primers / Q000235:genetics; D010505:Familial Mediterranean Fever / Q000175:diagnosis* / Q000235:genetics*; D005787:Gene Frequency; D005820:Genetic Testing / Q000379:methods; D005838:Genotype; D006579:Heterozygote; D006720:Homozygote; D006801:Humans; D009154:Mutation; D009693:Nucleic Acid Hybridization / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D000071198:Pyrin
chemical_list: D003598:Cytoskeletal Proteins; D017931:DNA Primers; C000604192:MEFV protein, human; D000071198:Pyrin