title: Detection of DeltaF508del using quantitative real-time PCR, comparison of the results obtained by fluorescent PCR.
abstract: OBJECTIVE
  Cystic fibrosis (CF) is the most common autosomal recessive genetic disorder in the Caucasian population. The molecular diagnosis is difficult since there are about 1,000 mutations in the CF transmembrane regulator gene. The DeltaF508del is the cause of the CF in 64% of the cases in Hungary. Our aim was to compare two polymerase chain reaction (PCR)-based method for the detection of DeltaF508del.
  
  
  METHODS
  One hundred and sixteen DNA samples isolated from different tissues (84 blood samples, 18 chorionic villus samples and 14 amniotic fluid samples) were involved in the study. Fluorescent PCR with DNA fragment analysis and quantitative real-time PCR with melting curve analysis were performed on the DNA samples for the detection of DeltaF508del.
  
  
  RESULTS
  Sixty-five healthy normal samples, 43 heterozygous samples, 6 DeltaF508del homozygous samples and 2 DeltaF508C homozygous samples were detected by using quantitative real-time PCR combined with melting curve analysis. The fluorescent PCR method did not detect the DeltaF508C mutation and these two samples were diagnosed as normal healthy ones.
  
  
  CONCLUSIONS
  The quantitative real-time PCR with melting curve analysis is a reliable and fast method for the detection of DeltaF508del. The results are ready in 1 h following the DNA isolation. The applied primer-probe set with melting curve analysis gives additional information for the presence of other mutations in the DeltaF508del region.
authors.affiliation: 1st Department of Obstetrics and Gynecology, Semmelweis University, Budapest, Hungary. nabal@noi1.sote.hu
authors.forename: Bálint
authors.initials: B
authors.lastname: Nagy
authors.forename: Gyula Richárd
authors.initials: GR
authors.lastname: Nagy
authors.forename: Levente
authors.initials: L
authors.lastname: Lázár
authors.forename: Zoltán
authors.initials: Z
authors.lastname: Bán
authors.forename: Zoltán
authors.initials: Z
authors.lastname: Papp
mesh_terms: D003550:Cystic Fibrosis / Q000175:diagnosis* / Q000235:genetics; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics*; D005260:Female; D006801:Humans; D009154:Mutation; D016133:Polymerase Chain Reaction / Q000379:methods*; D011247:Pregnancy; D011296:Prenatal Diagnosis / Q000379:methods*
chemical_list: D019005:Cystic Fibrosis Transmembrane Conductance Regulator