DNA analysis of cystic fibrosis in Brazil by direct PCR amplification from Guthrie cards

• Published in: American journal of medical genetics Vol. 46; no. 6; p. 665
• Main Authors: Raskin, S, Phillips, 3rd, J A, Krishnamani, M R, Vnencak-Jones, C, Parker, R A, Rozov, T, Cardieri, J M, Marostica, P, Abreu, F, Giugliani, R
• Format: Journal Article
• Language: English
• Published: United States 01.07.1993
• Subjects: Base Sequence; Brazil - epidemiology; Child; Cystic Fibrosis - diagnosis; Cystic Fibrosis - epidemiology; Cystic Fibrosis - genetics; DNA; DNA Mutational Analysis; Female; Gene Frequency; Humans; Male; Molecular Sequence Data; Mutation; Polymerase Chain Reaction; Brazil
• ISSN: 0148-7299
• DOI: 10.1002/ajmg.1320460612

A 3 bp deletion of codon 508 (phenylalanine) of the cystic fibrosis (CF) gene constitutes the mutation of most CF chromosomes. The frequency of this mutation (referred to as delta F508), varies considerably between populations, ranging from 26% of the CF mutations in Turkey to 88% in Denmark. To determine the frequency of the delta F508 mutation in Brazilian Caucasoid CF patients, we used direct polymerase chain reaction (PCR) amplification of DNA obtained from dried blood spots on Guthrie cards, followed by ethidium bromide staining of gels. Although the overall frequency of the delta F508 mutation was 47% of 380 CF chromosomes from Brazilian Caucasoids born in five different states, significant interstate differences were observed, ranging from a delta F508 frequency of 27% to 53%. While our method could be used to screen patients and their relatives for carrier testing and prenatal diagnosis, the efficacy of screening only for the delta F508 mutation would be low, and would vary from state to state. Screening for a panel of local mutations will be needed to increase the mutation detection rate and optimize genetic counseling.