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Detection of multiple cystic fibrosis mutations by reverse dot blot hybridization: a technology for carrier screening


We describe the implementation of a modified version of the reverse dot blot hybridization technology to detect eight cystic fibrosis mutations. The method is simple, quick, reliable, inexpensive, and nonradioactive and utilizes the sensitivity of the polymerase chain reaction coupled with colored or chemiluminescent substrates for mutation detection. We have used this system in a clinical laboratory to identify the ΔF508, G542X, G551D, R553X, 621 + 1G→T, W1282X, N1303K, and 1717G→A mutations. The technique is practical for genotyping individuals at many potential mutation sites, as in cystic fibrosis and β-thalassemia, in which over 95 mutations can cause disease. This technology appears to be the method of choice for the widespread carrier screening of multiple cystic fibrosis mutations.

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  1. Chehab FF, Doherty M, Cai SP, Kan YW, Cooper S, Rubin EM (1987) Detection of sickle cell anemia and thalassemias. Nature 329:293–294

  2. Chirgwin JM, Przybyla AE, MacDonald RJ, Rutter WJ (1979) Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry 18:5294–5296

  3. Cotton RG, Rodrigues 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

  4. Cutting GR, Kasch LM, Rosenstein BJ, Zielensky J, Tsui LC, Antonarakis SE, Kazazian HH Jr (1990) A cluster of cystic fibrosis mutations in the first nucleotide-binding fold of the cystic fibrosis conductance regulator protein. Nature 346:366–369

  5. Dean M, White MG, Amos J, Gerrard B, Stewart C, Khaw KT, Leppert M (1990) Multiple mutations in highly conserved residues are found in mildly affected cystic fibrosis patients. Cell 61:863–870

  6. Fries JW, Patel RJ, Piessens WF, Wirth DF (1990) Genus and species-specific DNA probes to identify mycobacteria using the polymerase chain reaction. Mol Cell Probes 2:87–105

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

  8. Hance AJ, Grandchamp B, Levy-Frebault V, Lecossier D, Rauzier J, Bocart D, Gicquel B (1989) Detection and identification of mycobacteria by amplification of mycobacterial DNA. Mol Microbiol 3:843–849

  9. Kawasaki ES (1990) Sample preparation from blood, cells and other fluids. In: Innis MA, Gelfand DH, Sninsky JJ, White TJ (eds) PCR protocols. Academic Press, New York

  10. 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

  11. Kerem BS, Zielenski J, Markiewicz D, Bozon D, Gazi E, Yahav J, Kennedy D, Riordan JR, Collins FS, Rommens JM, 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 87:8847–8851

  12. Klein D, Dodson AE, Tabor D, Cederbaum SD, Grody WW (1991) Effect of an adjacent base on detection of a point mutation by restriction enzyme digestion. Somat Cell Mol Genet 17:369–375

  13. 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

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

  15. 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 J, Tsui LC, Collins FS (1989) Identification of the cystic fibrosis gene: chromosome walking and jumping. Science 245:1059–1065

  16. Saiki RS, Gelfand DH, Stoffel S, Scharf SJ, Higuchi R, Horn GT, Mullis KB, Erlich HA (1988) Primer directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 239:487–491

  17. Saiki RK, Walsh PS, Levenson CH, Erlich HA (1989) Genetic analysis of amplified DNA with immobilized sequence-specific oligonucleotide probes. Proc Natl Acad Sci USA 86:6230–6234

  18. 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

  19. Watson E, Williamson B, Brueton L, Winter R (1990) Genetic counselling for cystic fibrosis based upon mutation/haplotype analysis. Lancet 336:190–191

  20. Zhang Y, Coyne MY, Will SG, Levenson CH, Kawasaki ES (1991) Single base mutational analysis of cancer and genetic diseases using membrane bound modified oligonucleotides. Nucleic Acids Res 19:3929–3933

  21. Zielenski J, Rozmahel R, Bozon D, Kerem BS, Grelczack Z, Riordan Jr, Rommens J, Tsui LC (1991a) Genomic DNA sequence of the cystic fibrosis transmembrane conductance regulator (CFTR) gene. Genomics 10:214–228

  22. Zielenski J, Bozon D, Kerem B, Markiewicz D, Rommens JM, Tsui LC (1991b) Identifications of mutations in exons 1 through 8 of the cystic fibrosis transmembrane conductance regulator (CFTR) gene. Genomics 10:229–235

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Chehab, F.F., Wall, J. Detection of multiple cystic fibrosis mutations by reverse dot blot hybridization: a technology for carrier screening. Hum Genet 89, 163–168 (1992). https://doi.org/10.1007/BF00217117

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  • Polymerase Chain Reaction
  • Internal Medicine
  • Cystic Fibrosis
  • Metabolic Disease
  • Clinical Laboratory