Skip to main content
PMC home page
Journal of Medical Genetics logoLink to Journal of Medical Genetics
. 1991 May;28(5):304–311. doi: 10.1136/jmg.28.5.304

A convenient multiplex PCR system for the detection of dystrophin gene deletions: a comparative analysis with cDNA hybridisation shows mistypings by both methods.

S Abbs 1, S C Yau 1, S Clark 1, C G Mathew 1, M Bobrow 1
PMCID: PMC1016847  PMID: 1865467

Abstract

Existing reactions for the multiplex PCR amplification of exons in the dystrophin gene have been modified to produce two multiplex reactions which separately cover the 5' and 3' major deletion 'hotspots' in the gene, and together detect approximately 98% of all deletions detectable by Southern cDNA hybridisation. A comparative study of 148 patients showed mistypings in both the cDNA hybridisation data (4%) and the PCR analysis (1.2%). We suggest means of circumventing the underlying problems in order to avoid mistyping and subsequent misdiagnosis, and conclude that, with appropriate precautions, multiplex PCR amplification can be the method of choice for detecting deletions in the dystrophin gene.

Full text

PDF
304

Images in this article

305Image
on p.305
307Image
on p.307
309Image
on p.309

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abbs S., Roberts R. G., Mathew C. G., Bentley D. R., Bobrow M. Accurate assessment of intragenic recombination frequency within the Duchenne muscular dystrophy gene. Genomics. 1990 Aug;7(4):602–606. doi: 10.1016/0888-7543(90)90205-9. [DOI] [PubMed] [Google Scholar]
  2. Beggs A. H., Koenig M., Boyce F. M., Kunkel L. M. Detection of 98% of DMD/BMD gene deletions by polymerase chain reaction. Hum Genet. 1990 Nov;86(1):45–48. doi: 10.1007/BF00205170. [DOI] [PubMed] [Google Scholar]
  3. Burghes A. H., Logan C., Hu X., Belfall B., Worton R. G., Ray P. N. A cDNA clone from the Duchenne/Becker muscular dystrophy gene. 1987 Jul 30-Aug 5Nature. 328(6129):434–437. doi: 10.1038/328434a0. [DOI] [PubMed] [Google Scholar]
  4. Chamberlain J. S., Gibbs R. A., Ranier J. E., Nguyen P. N., Caskey C. T. Deletion screening of the Duchenne muscular dystrophy locus via multiplex DNA amplification. Nucleic Acids Res. 1988 Dec 9;16(23):11141–11156. doi: 10.1093/nar/16.23.11141. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Fujimura F. K., Northrup H., Beaudet A. L., O'Brien W. E. Genotyping errors with the polymerase chain reaction. N Engl J Med. 1990 Jan 4;322(1):61–61. doi: 10.1056/NEJM199001043220117. [DOI] [PubMed] [Google Scholar]
  6. Green P. M., Bentley D. R., Mibashan R. S., Nilsson I. M., Giannelli F. Molecular pathology of haemophilia B. EMBO J. 1989 Apr;8(4):1067–1072. doi: 10.1002/j.1460-2075.1989.tb03474.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hodgson S. V., Bobrow M. Carrier detection and prenatal diagnosis in Duchenne and Becker muscular dystrophy. Br Med Bull. 1989 Jul;45(3):719–744. doi: 10.1093/oxfordjournals.bmb.a072354. [DOI] [PubMed] [Google Scholar]
  8. Hodgson S., Hart K., Abbs S., Heckmatt J., Rodillo E., Bobrow M., Dubowitz V. Correlation of clinical and deletion data in Duchenne and Becker muscular dystrophy. J Med Genet. 1989 Nov;26(11):682–693. doi: 10.1136/jmg.26.11.682. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hu X. Y., Ray P. N., Murphy E. G., Thompson M. W., Worton R. G. Duplicational mutation at the Duchenne muscular dystrophy locus: its frequency, distribution, origin, and phenotypegenotype correlation. Am J Hum Genet. 1990 Apr;46(4):682–695. [PMC free article] [PubMed] [Google Scholar]
  10. Koenig M., Beggs A. H., Moyer M., Scherpf S., Heindrich K., Bettecken T., Meng G., Müller C. R., Lindlöf M., Kaariainen H. The molecular basis for Duchenne versus Becker muscular dystrophy: correlation of severity with type of deletion. Am J Hum Genet. 1989 Oct;45(4):498–506. [PMC free article] [PubMed] [Google Scholar]
  11. Koenig M., Hoffman E. P., Bertelson C. J., Monaco A. P., Feener C., Kunkel L. M. Complete cloning of the Duchenne muscular dystrophy (DMD) cDNA and preliminary genomic organization of the DMD gene in normal and affected individuals. Cell. 1987 Jul 31;50(3):509–517. doi: 10.1016/0092-8674(87)90504-6. [DOI] [PubMed] [Google Scholar]
  12. Koenig M., Monaco A. P., Kunkel L. M. The complete sequence of dystrophin predicts a rod-shaped cytoskeletal protein. Cell. 1988 Apr 22;53(2):219–228. doi: 10.1016/0092-8674(88)90383-2. [DOI] [PubMed] [Google Scholar]
  13. Malhotra S. B., Hart K. A., Klamut H. J., Thomas N. S., Bodrug S. E., Burghes A. H., Bobrow M., Harper P. S., Thompson M. W., Ray P. N. Frame-shift deletions in patients with Duchenne and Becker muscular dystrophy. Science. 1988 Nov 4;242(4879):755–759. doi: 10.1126/science.3055295. [DOI] [PubMed] [Google Scholar]
  14. Monaco A. P., Bertelson C. J., Liechti-Gallati S., Moser H., Kunkel L. M. An explanation for the phenotypic differences between patients bearing partial deletions of the DMD locus. Genomics. 1988 Jan;2(1):90–95. doi: 10.1016/0888-7543(88)90113-9. [DOI] [PubMed] [Google Scholar]
  15. Prior T. W., Friedman K. J., Silverman L. M. RFLP for HindIII at the Duchenne muscular dystrophy gene. Nucleic Acids Res. 1989 Mar 25;17(6):2370–2370. doi: 10.1093/nar/17.6.2370. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Roberts R. G., Cole C. G., Hart K. A., Bobrow M., Bentley D. R. Rapid carrier and prenatal diagnosis of Duchenne and Becker muscular dystrophy. Nucleic Acids Res. 1989 Jan 25;17(2):811–811. doi: 10.1093/nar/17.2.811. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Saiki R. K., Scharf S., Faloona F., Mullis K. B., Horn G. T., Erlich H. A., Arnheim N. Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. Science. 1985 Dec 20;230(4732):1350–1354. doi: 10.1126/science.2999980. [DOI] [PubMed] [Google Scholar]
  18. Winship P. R. An improved method for directly sequencing PCR amplified material using dimethyl sulphoxide. Nucleic Acids Res. 1989 Feb 11;17(3):1266–1266. doi: 10.1093/nar/17.3.1266. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Medical Genetics are provided here courtesy of BMJ Publishing Group

RESOURCES