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Human Genetics

, Volume 86, Issue 1, pp 45–48 | Cite as

Detection of 98% of DMD/BMD gene deletions by polymerase chain reaction

  • Alan H. Beggs
  • Michel Koenig
  • Frederick M. Boyce
  • Louis M. Kunkel
Original Investigations

Summary

We describe oligonucleotide primer sequences that can be used to amplify eight exons plus the muscle promoter of the dystrophin gene in a single multiplex polymerase chain reaction (PCR). When used in conjunction with an existing primer set, these two multiplex reactions detect about 98% of deletions in patients with Duchenne or Becker muscular dystrophy (DMD, BMD). Furthermore, these primers amplify most of the exons in the deletion prone “hot spot” region around exons 44 to 53, allowing determination of deletion endpoints and prediction of mutational effects on the translational reading frame. Thus, use of these PCR-based assays will allow deletion detection and prenatal diagnosis for most DMD/BMD patients in a fraction of the time required for Southern blot analysis.

Keywords

Southern Blot Muscular Dystrophy Southern Blot Analysis Oligonucleotide Prime Prenatal Diagnosis 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. Baumbach LL, Chamberlain JS, Ward PA, Farwell NJ, Caskey CT (1989) Molecular and clinical correlations of deletions leading to Duchenne and Beeker muscular dystrophies. Neurology 39:465–474Google Scholar
  2. Beggs AH, Kunkel LM (1990a) Improved diagnosis of Duchenne/ Becker muscular dystrophy. J Clin Invest 85:613–619Google Scholar
  3. Beggs AH, Kunkel LM (1990b) A polymorphic CACA repeat in the 3′ untranslated region of dystrophin. Nucleic Acids Res 18:1931Google Scholar
  4. Chamberlain JS, Gibbs RA, Rallier JE, Nguyen PN, Caskey CT (1988) Deletion screening of the Duchenne muscular dystrophy locus via multiplex DNA amplification. Nucleic Acids Res 16:11141–11156Google Scholar
  5. Chamberlain JS, Gibbs RA, Ranier JE, Caskey CT (1990) Multiplex PCR for the diagnosis of Duchenne muscular dystrophy. In: Innis MA, Gelfand DH, Sninsky JJ, White TJ (eds) PCR protocols: a guide to methods and applications. Academic Press. New York London, pp 272–281Google Scholar
  6. Forrest SM, Cross GS, Flint T, Speer A, Robson KJH, Davies KE, (1988) Further studies of gene deletions that cause Duchenne and Becker muscular dystrophies. Genomics 2:109–114Google Scholar
  7. Gillard EF, Chamberlain JS, Murphy EG, Duff CL, Smith B, Burghes AHM, Thompson MW, Sutherland J, Oss I, Bodrug SE, Klamut HJ, Ray PN, Worton RG (1989) Molecular and phenotypic analysis of patients with deletions within the deletion-rich region of the Duchenne muscular dystrophy (DMD) gene. Am J Hum Genet 45:507–520Google Scholar
  8. Hoffman EP, Kunkel LM (1989) Dystrophin abnormalities in Duchenne/Becker muscular dystrophy. Neuron 2:1019–1029Google Scholar
  9. Hoffman EP, Fischbeek KH, Brown RH, Johnson M, Medori R, Loike JD, Harris JB, Waterston R, Brookc M, Specht L, Kupsky W, Chamberlain J, Caskey CT, Shapiro F, Kunkel LM (1988) Characterization of dystrophin in muscle-biopsy specimens from patients with Duchenne's or Seeker's muscular dystrophy. N Engl J Med 318:1363–1368Google Scholar
  10. Hoffman EP, Kunkel LM, Angelini C, Clarke A, Johnson M, Harris JB (1989a) Improved diagnosis of Becker muscular dystrophy via dystrophin testing. Neurology 39:1011–1017Google Scholar
  11. Hoffman EP, Beggs AH, Koenig M, Kunkel LM, Angelini C (1989b) Cross-reactive protein in Duchenne muscle. Lancet 11:1211–1212Google Scholar
  12. Hu X, Burghes AHM, Ray PN, Thompson MW, Murphy EG, Worton RG (1988) Partial gene duplication in Duchenne and Becker muscular dystrophy. J Med Genet 25:369–376Google Scholar
  13. Innis MA, Gelfand DH, Sninsky JJ, White TJ (eds) (1990) PCR protocols: a guide to methods and applications. Academic Press. New York LondonGoogle Scholar
  14. Kawasaki ES (1990) Sample preparation from blood, cells, and other fluids. In: Innis MA, Gelfand DH, Sninsky JJ, White TJ (eds) PCR protocols: a guide to methods and applications. Academic Press, New York London, pp 146–152Google Scholar
  15. Koenig M, Hoffman EP, Bertelson CJ, Monaco AP, Feener C, Kunkel LM (1987) Complete cloning of the Duchenne muscular dystrophy (DMD) cDNA and preliminary genomic organization of the DMD gene in normal and affected individuals. Cell 50:509–517Google Scholar
  16. Koenig M, Monaco AP, Kunkel LM (1988) The complete sequence of dystrophin predicts a rod-shaped cytoskeletal protein. Cell 53:219–228Google Scholar
  17. Koenig M, Beggs AH, Moyer M, Scherpf S, Heindrichs K, Bettecken T, Meng G, Muller CR, Lindlof M, Kaariainen H, Chapelle A de la, Kiuru A, Savontaus M-L, Gilgenkrantz H, Recan D, Chelly J, Kaplan J-C, Covonc AE, Archidiacono N, Romeo G, Liechti-Gallati S, Schneider V, Braga S, Moser H, Darras BT, Murphy P, Franeke U, Chen JD, Morgan G, Denton M, Greeneberg CR, Wrogernann K, Blondon LAJ, Paassen HMB van, Ommen GJB van, Kunkel LM (1989) The molecular basis for Duchenne versus Becker muscular dystrophy: correlation of severity with type of deletion. Am J Hum Genet 45:498–506PubMedGoogle Scholar
  18. Kunkel LM, Smith KD, Boyer SH, Borgaonkar DS, Wachtel SS, Miller OJ, Breg WR, Jones HW, Rary JM (1977) Analysis of human Y-chromosome-specific reiterated DNA in chromosome variants. Proc Natl Acad Sci USA 74:1245–1249PubMedGoogle Scholar
  19. Malhotra SB, Hart KA, Klamut HJ, Thomas NST, Bodrug SE, Burghes AHM, Bobrow M, Harper PS, Thompson MW, Ray PN, Worton RG (1988) Frame-shift deletions in patients with Duchenne and Beeker muscular dystrophy. Science 242:755–759Google Scholar
  20. Monaco AP, Bertelson CJ, Liechti-Gallati S, Moser H, Kunkel M (1988) An explanation for the phenotypic differences between patients bearing partial deletions of the DMD locus. Genomics 2:90–95Google Scholar
  21. Roberts RG, Cole CG, Hart KA, Bobrow M, Bentley DR (1989) Rapid carrier and prenatal diagnosis of Duchenne and Becker muscular dystrophy. Nucleic Acids Res 17:811Google Scholar

Copyright information

© Springer-Verlag 1990

Authors and Affiliations

  • Alan H. Beggs
    • 1
    • 2
  • Michel Koenig
    • 1
    • 2
  • Frederick M. Boyce
    • 1
    • 2
  • Louis M. Kunkel
    • 1
    • 2
  1. 1.Genetics Division, Children's HospitalHoward Hughes Medical InstituteBostonUSA
  2. 2.Department of PediatricsHarvard Medical SchoolBostonUSA

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