title: Non-isotopic RNase cleavage assay for mutation detection in MEFV, the gene responsible for familial Mediterranean fever, in a cohort of Greek patients.
abstract: BACKGROUND
  The MEFV gene is responsible for familial Mediterranean fever (FMF). Several disease associated mutations have been identified. The range of genetic variation in MEFV in Greek patients has not been determined.
  
  
  OBJECTIVE
  To describe a method that facilitates the routine screening of the entire coding sequence of MEFV (excluding exon 1).
  
  
  METHODS
  The non-isotopic RNase cleavage assay (NIRCA) was optimised and used as a first step screening method to screen exons 2 to 10 of MEFV. Exons 2 and 10 were analysed separately at DNA level, while exons 3 to 9 were analysed together at cDNA level. The sample group consisted of 26 FMF patients diagnosed using established clinical criteria, six asymptomatic relatives, 12 patients with atypical clinical manifestations, nine patients suffering from various inflammatory diseases, and three normal individuals. All were analysed by NIRCA for mutations in the MEFV gene and direct sequencing was applied subsequently to confirm the results.
  
  
  RESULTS
  MEFV mutations were identified in 25 of 26 typical FMF patients and in two of 12 patients with atypical manifestations. NIRCA results were in concordance with sequencing findings in all sequences analysed, suggesting that the method is highly reliable in this disease. Sixteen alterations of MEFV were identified (eight missense mutations and eight single nucleotide polymorphisms).
  
  
  CONCLUSIONS
  NIRCA can be used for rapid screening of the coding sequence of the MEFV gene in patients suspected of suffering from FMF.
authors.affiliation: First Division of Internal Medicine, Democritus University of Thrace, Alexandroupolis, Greece. ritis2@otenet.gr
authors.forename: K
authors.initials: K
authors.lastname: Ritis
authors.forename: S
authors.initials: S
authors.lastname: Giaglis
authors.forename: N
authors.initials: N
authors.lastname: Spathari
authors.forename: A
authors.initials: A
authors.lastname: Micheli
authors.forename: D
authors.initials: D
authors.lastname: Zonios
authors.forename: D
authors.initials: D
authors.lastname: Tzoanopoulos
authors.forename: C C
authors.initials: CC
authors.lastname: Deltas
authors.forename: S
authors.initials: S
authors.lastname: Rafail
authors.forename: R
authors.initials: R
authors.lastname: Mean
authors.forename: V
authors.initials: V
authors.lastname: Papadopoulos
authors.forename: A G
authors.initials: AG
authors.lastname: Tzioufas
authors.forename: H M
authors.initials: HM
authors.lastname: Moutsopoulos
authors.forename: G
authors.initials: G
authors.lastname: Kartalis
mesh_terms: D000293:Adolescent; D000328:Adult; D001483:Base Sequence; D002675:Child, Preschool; D015331:Cohort Studies; D003598:Cytoskeletal Proteins; D010505:Familial Mediterranean Fever / Q000453:epidemiology / Q000235:genetics*; D005260:Female; D006115:Greece / Q000453:epidemiology; D006720:Homozygote; D006801:Humans; D008297:Male; D008875:Middle Aged; D009154:Mutation / Q000235:genetics; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction / Q000379:methods; D011506:Proteins / Q000235:genetics*; D000071198:Pyrin; D012260:Ribonucleases / Q000378:metabolism*
chemical_list: D003598:Cytoskeletal Proteins; C000604192:MEFV protein, human; D011506:Proteins; D000071198:Pyrin; D012260:Ribonucleases