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Retrospective study of the cystic fibrosis transmembrane conductance regulator (CFTR) gene mutations in Guthrie cards from a large cohort of neonatal screening for cystic fibrosis

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Abstract

The cystic fibrosis transmembrane conductance regulator (CFTR) gene encodes a cAMP-activated chloride channel, and in individuals with both alleles of the gene mutated, symptoms of CF disease are manifest. With more than 300 mutations so far described in the gene the profile of mutant alleles in a population is specific to its ethnic origin. For an analysis with an unbiased recruitment of the CF alleles in neonates of similar origin (Normandy, France), we have retrospectively analyzed the Guthrie cards of affected newborns, diagnosed by the immunoreactive trypsinogen (IRT) assay. Analysis of the 27 exons of the CFTR gene using a GC clamp denaturing gradient gel electrophoresis (DGGE) assay has enabled us to identify over 96% of the mutated alleles. Two of these were novel mutations. We would like to propose this strategy as an efficient method of retrospective molecular genetic diagnosis that can be performed wherever Guthrie cards can be obtained. Knowledge of rare alleles could be a prerequisite for CF therapy in the future.

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References

  • Abeliovich D, Lavon IP, Lerer I, Cohen T, Springer C, Avital A, Cutting GR (1992) Screening for five mutations detects 97% of cystic fibrosis (CF) chromosomes and predicts a carrier frequency of 1∶29 in the Jewish Ashkenazi population. Am J Hum Genet 51:951–956

    Google Scholar 

  • Aubin G, et al. (1992) Cystic fibrosis mutations in French Canadians: three CFTR mutations are relatively frequent in a Quebec population with an elevated incidence of cystic fibrosis. Am J Med Genet 42:360–364

    Google Scholar 

  • Audrézet MP, Mercier B, Guillermit H, Quéré I, Verlingue C, Rault G, Férec C (1993) Identification of 12 novel mutations in the CFTR gene. Hum Mol Genet 2:51–54

    Google Scholar 

  • Audrézet MP, Costes B, Ghanem N, Fanen P, Verlingue C, Morin JF, Mercier B, Goossens M, Férec C (1993) Screening for CF in dried blood spots of newborns. Mol and Cell Probes (in press)

  • Bear EC, Li C, Karner N, Bridges RJ, Jensen TJ, Ramjeesingh M, Riordan JR (1992) Purification and functional reconstitution of the ccystic fibrosis transmembrane conductance regulator (CFTR). Cell 68:809–818

    Article  CAS  PubMed  Google Scholar 

  • Boat TF, Welsh MJ, Beaudet AL (1989) In: Scriver CR, Beaudet AL, Sly WS, Valle D (eds) The metabolic basis of inherited disease. McGraw Hill, New-York, pp 2649–2680

    Google Scholar 

  • Claustres M, Desgeorges M, Kjellberg P, Bellet H, Demaille J, Ramsay M (1990) Cystic fibrosis typing with DNA probes and screening for delta F508 deletion in families from Southern France. Hum Genet 85:398–399

    Google Scholar 

  • Claustres M, Gerrard B, White MB, Desgeorges M, Kjellberg P, Rollin B, Dean M (1992) A rare mutation (1078 del T) in exon 7 of the CFTR gene in a Southern French adult with cystic fibrosis. Genomics 13:907

    Google Scholar 

  • Claustres M, Maguelone L, Desgeorges M, Giansily M, Culard JF, Razakatsara G, Gerrard B, Demaille J (1993) Analysis of the 27 exons and flanking regions of the cystic fibrosis gene: 40 different mutaions account for 91.2% of the mutant alleles in Southern France. Hum Mol Genet 2:1209–1213

    Google Scholar 

  • Costes B, Fanen P, Goossens M, Ghanem N (1993) A rapid, efficient and sensitive assay for simultaneous detection of multiple cystic fibrosis mutations. Hum Mutat 2:185–191

    Google Scholar 

  • Cotton RGH, Rodriquez NR, Campbell RD (1988) Reactivity of cytosine and thymine in single-base-pair mismatches with hydroxylamine and osmium tetroxide and its application to the study of mutations. Proc Natl Acad Sci USA 85:4397–4401

    Google Scholar 

  • Cremonesi L, Ferrari M, Belloni E, Magnani C, Seia M, Ronchetto P, Rady M, Russo MP, Romeo G, Devoto M. (1992) Four new mutations in the CFTR gene (541delC, R347H, R352Q, E585X) detected by DGGE analysis in Italian CF patients, associated with different clinical phenotypes. Hum Mutats 1:314–319

    Google Scholar 

  • Cutting GR, Kasch LM, Rosenstein GJ, Zielenski J, Tsui LC, Kazazian Jr HH, Antonarakis SE (1990) Two patients with cystic fibrosis, non-sense mutations in each cystic fibrosis gene, and mild pulmonary disease. N Engl J Med 323:1685–1689

    Google Scholar 

  • Dalemans W, Barbry P, Champigny G, Jallat S, Dott K, Dreyer D, Crystal RG, Pavirani A, Lecocq JP, Lazdunski M (1991) Altered chloride ion channel kinetics associated with the ΔF508 cystic fibrosis mutation. Nature 354:526–528

    Google Scholar 

  • Fanen P, Ghanem N, Vidaud M, Besmond C, Martin J, Costes P, Plassa F, Goossen M (1992) Molecular characterization of cystic fibrosis: 16 novel mutations identified by analysis of the whole cystic fibrosis transmembrane conductance regulator (CFTR) coding regions and splice sites. Genomics 13:770–776

    Google Scholar 

  • Férec C, Audrézet MP, Mercier B, Guillermit H, Moulllier P, Quéré I, Verlingue C (1992) Detection of over 98% of cystic fibrosis mutations in a Celtic population. Nature Genet 1:188–191

    Google Scholar 

  • Férec C, Mercier B, Verlingue C, Guillermit H, Audrézet MP, Feigelson J (1993) Genotype/phenotype correlation in adult healthy cystic fibrosis patients. 25th Annual Meeting of the European Society of Human Genetics, Barcelona, 6–9 May

  • Gasparini P, Nunes V, Savoia A, Dognini M, Morral N, Gaona A, Bonizzato A, Chillon M, Sangiuolo F, Novelli G, Dallapiccola B, Pignatti PF, Estivill X (1991) The search for South European cystic fibrosis mutations: identification of two new mutations, four variants, and intronic sequences. Genomics 10:193–200

    Google Scholar 

  • Gregg GR, Wilfond BS, Farrell PM, Laxova A, Hassemer D, Mischler EH (1993) Application of DNA analysis in a populationscreening program for neonatal diagnosis of cystic fibrosis (CF): comparison of screening protocols. Am J Hum Genet 52:616–626

    Google Scholar 

  • Guillermit H, Fanen P, Férec C (1990) A 3′ splice consensus sequence mutation in the cystic fibrosis gene. Hum Genet 85:450–453

    Google Scholar 

  • Gyllensten UB, Erlich HA (1988) Generation of single-stranded DNA by the polymerase chain reaction and its application to direct sequencing of the HLA-DQA locus. Proc Natl Acad Sci USA 85:7652–7656

    Google Scholar 

  • Hammond KB, Reardon MC, Accurso FJ, Abman SH, Cotton EK, Pinney M, Fisher C, et al. (1986) Early detection and follow-up of cystic fibrosis in newborns: the Colorado experience In: Carter TP, Willey AM (eds) Genetic disease: screening and management. Liss, New York, pp 81–101

    Google Scholar 

  • Hammond KB, Abman SH, Sokol RJ, Accurso FJ (1991) Efficacy of statewide neonatal screening for cystic fibrosis by assay of trypsinogen concentrations. N Engl J Med 325:769–774

    Google Scholar 

  • Kerem BS, Rommens JM, Buchanan JA, Markiewicz D, Cox TK, Chakravarti A, Buchwald M, Tsui LC (1989) Identification of the cystic fibrosis gene: genetic analysis. Science 245:1073–1080

    Google Scholar 

  • Kerem B, Zielenski J, Markiewicz D, Bozon D, Gazit E, Yaha, J, Kennedy D, Riordan JR, Collins FS, Rommens JR, Tsui LC (1990) Identification of mutations in regions corresponding to the 2 putative nucleotide (ATP) binding folds of the cystic fibrosis gene. Proc Natl Acad Sci USA 87:8447–8451

    Google Scholar 

  • Kristidis P, Bozon D, Corey M, Markiewicz D, Rommens J, Tsui LC, Durie P (1992) Genetic determination of exocrine pancreatic function in cystic fibrosis. Am J Hum Genet 50:1178–1184

    Google Scholar 

  • Laroche D, Travert G (1991) The application of PCR amplification and the polymorphic marker KM19 to dried blood spots: comparison with deletion 508 for the confirmation of the neonatal screening test for cystic fibrosis. Pediatr Pulmonol [Suppl] 7:19–22

    Google Scholar 

  • Lerman LS, Silverstein K (1987) Computational simulation of DNA melting and its application to denaturing gradient gel electrophoresis. Methods Enzymol 155:482–501

    Google Scholar 

  • Mercier B, Lissens W, Novelli G, Kalaydjieva L, De Arce M, Kapranov N, Canki Klain N, Lenoir G, Chauveau P, Lenaerts C, Cashman S, Sangiuolo F, Audrézet MP, Dallapiccola B, Guillermit H, Bonduelle M, Liebaers I, Quéré I, Verlingue C, Férec C (1993) Identification of eight novel mutations in a collaborative analysis of a part of the second transmembrane domain of the CFTR gene. Genomics 16:296–297

    Google Scholar 

  • Morral N, Nunes V, Casals T, Chillon M, Giménez J, Bertranpetit J, Estivill X (1993) Microsatellite haplotypes for cystic fibrosis: mutation frameworks and evolutionary tracers. Hum Mol Genet 2:1015–1022

    Google Scholar 

  • Myers RM, Maniatis T, Lerman LS (1987) Detection and localization of single base changes by denaturing gradient gel electrophoresis. Methods Enzymol 155:501–527

    Google Scholar 

  • Nunes V, Chillon M, Dörk T, Tammler B, Casals T, Estivill X (1993) A new missense mutation (E92K) in the first transmembrane domain of the CFTR gene causes a benign cystic fibrosis phenotype. Hum Mol Genet 2:79–80

    Google Scholar 

  • Orita M, Suzuki Y, Sekiya T, Hayashi K (1989) Rapid and sensitive detection of point mutations and DNA polymorphisms using the polymerase chain reaction. Genomics 5:874–879

    CAS  PubMed  Google Scholar 

  • Osborne L, Knight R, Santis G, Hodson M (1991) A mutation in the second nucleotide binding fold of the cystic fibrosis gene. Am J Hum Genet 48:608–612

    Google Scholar 

  • Riordan JR, Rommens JM, Kerem BS, Alon N, Rozmahel R, Grzelczak Z, Zielenski J, Lok S, Plavsic N, Chou JL, Drumm ML, Iannuzzi MC, Collins FS, Tsui LC (1989) Identification of the cystic fibrosis gene: cloning and characterization of complementary DNA. Science 245:1066–1073

    CAS  PubMed  Google Scholar 

  • Rommens JM, Iannuzzi MC, Kerem BS, Drumm ML, Melmer G, Dean M, Rozmahel R, Cole JL, Kennedy D, Hidaka N, Zsiga M, Buchwald M, Riordan JR, Tsui LC, Collins FS (1989) Identification of the cystic fibrosis gene: chromosome walking and jumping. Science 245:1059–1064

    Google Scholar 

  • Rommens JM, Dho S, Bear CE, Kartner N, Kennedy D, Riordan JR, Tsui LC, Foskett JK (1991) cAMP-inducible chloride conductance in mouse fibroblast lines stably expressing the human cystic fibrosis transmembrane conductance regulator. Proc Natl Acad Sci USA 88:7500–7504

    Google Scholar 

  • Rozen R, Schwartz RH, Hilman BC, Stanislovitis P, Horn GT, Klinger K, Daigneault J, Braekeleer M de, Kerem BS, Tsui LC, Fujiwara TM, Morgan K (1992) Cystic fibrosis mutations in North American populations of French ancestry: analysis of Quebec French-Canadian and Louisiana Acadian families. Am J Hum Genet 47:606–610

    Google Scholar 

  • Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467

    CAS  PubMed  Google Scholar 

  • Schwartz M, Summers C, Heptinstall L, Newton C, Markham A, Cain R, Super M (1991) A deletion mutation of the cystic fibrosis transmembrane conductance regulator (CFTR) locus: ΔI507. Adv Exp Med Biol 290:393–398

    Google Scholar 

  • Sheffield VC, Cox DR, Lerman SL, Myers RM (1989) Attachment of a 40-base pair G + C-rich sequence (GC-clamp) to genomic DNA fragments by the polymerase chain reaction results in improved detection of single-base changes. Proc Natl Acad Sci USA 86:232–236

    Google Scholar 

  • Sheppard DN, Rich DP, Ostedgaard LS, Gregory RJ, Smith AE, Welsh M (1993) Mutations in CFTR associated with mild-disease-form Cl− channels with altered pore properties. Nature 362:160–164

    Google Scholar 

  • Shoshani T, Augarten A, Gazit E, Bashan N, Yahav Y, Rivlin Y, Tal A, Seret H, Yaar L, Kerem E, Kerem BS (1992) Association of a nonsense mutation (W1282X), the most common mutation in the Ashkenazi Jewish cystic fibrosis patients in Israel, with presentation of severe disease. Am J Hum Genet 50:222–228

    Google Scholar 

  • Travert G, Duhamel JF (1983) Neonatal screening for cystic fibrosis using trypsin radioimmunoassay in blood. Arch Fr Pediatr 40:295–298

    Google Scholar 

  • Tsui LC (1992) Mutations and sequence variations detected in the cystic fibrosis transmembrane conductance regulator (CFTR) gene: a report from the Cystic Fibrosis Genetic Analysis Consortium. Hum Mutat 1:197–203

    Google Scholar 

  • Tucker SJ, Tannahill D, Higgins CF (1992) Identification and developmental expression of the Xenopus laevis cystic fibrosis transmembrane conductance regulator gene. Hum Mol Gen 1:77–82

    Google Scholar 

  • Vidaud M, Fanen P, Martin J, Ghanem N, Nicolas S, Goossens M (1990) Three point mutations in the CFTR gene in French cystic fibrosis patients: identification by denaturing gradient gel electrophoresis. Hum Genet 85:446–449

    Google Scholar 

  • Zielenski J, Bozon D, Kerem BS, Markiewicz D, Durie P, Rommens JM, Tsui L.C. (1991) Identification of mutations in exons 1 through 8 of the cystic fibrosis transmembrane conductance regulator (CFTR) gene. Genomics 10:229–235

    Google Scholar 

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Authors and Affiliations

  1. Centre de Biogénétique, Centre Départemental de Transfusion Sanguine, 46 rue Félix le Dantec, BP 454, F-29275, Brest Cedex, France

    C. Verlingue, B. Mercier, M. P. Audrézet & C. Férec

  2. Laboratoire de Biophysique Médicale, CHU de Caen, Avenue de la Côte de Nacre, F-14033, Caen Cedex, France

    I. Lecoq, D. Laroche & G. Travert

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  1. C. Verlingue
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  2. B. Mercier
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  3. I. Lecoq
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  4. M. P. Audrézet
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  5. D. Laroche
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  6. G. Travert
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  7. C. Férec
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Verlingue, C., Mercier, B., Lecoq, I. et al. Retrospective study of the cystic fibrosis transmembrane conductance regulator (CFTR) gene mutations in Guthrie cards from a large cohort of neonatal screening for cystic fibrosis. Hum Genet 93, 429–434 (1994). https://doi.org/10.1007/BF00201669

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  • DOI: https://doi.org/10.1007/BF00201669

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