Summary
We compared the efficacy of the multiplex PCR with that of the cDNA analysis for detection of deletions of the DMD gene in the Japanese patients. Thirty males with DMD from 27 Japanese families were studied by the multiplex PCR, and 24 of them were also investigated by Southern blot analysis. We used five dystrophin cDNA probes for deletion analysis. A total of 19 regions were amplified by the PCR to detect deletions, 9 regions by the method of Chamberlainet al. and another 10 regions by the method of Beggset al. Deletions were detected in 14 (52%) out of 27 DMD families by the PCR. Southern blot analysis detected deletions in 14 (64%) out of 22 families. Thirteen (93%) of the 14 DMD families with deletions detected by Southern blotting were also confirmed by the multiplex PCR. Provided care is taken in cases where the deletion is limited to a single exon, the multiplex PCR appears to be an efficient and useful alternative to conventional Southern blot analysis for detecting deletions during the prenatal and postnatal diagnosis of DMD.
Similar content being viewed by others
Article PDF
References
Abbs S, Yau SC, Clark S, Mathew CG, Bobrow M (1991): A convenient multiplex PCR system for the detection of dystrophin deletion: A comparative analysis with cDNA hybridization reveals mistypings by both methods. J Med Genet28: 304–311
Asano J, Tomatsu S, Sukegawa K, Ikedo Y, Minami R, Iida M, Nishimura M, Nakagawa M, Ohsiro M, Orii T (1991): Gene deletions in the Japanese patients with Duchenne and Becker muscular dystrophies: Deletion study and carrier detection. Clin Genet39: 419–424
Beggs A, Koenig H, Boyce FM, Kunkel LM (1990): Detection of 98% of DMD/BMD gene deletions by polymerase chain reaction. Hum Genet86:45–48
Chamberlain JS, Gibbs RA, Ranier JF, Nguyen PN, Caskey CT (1988): Deletion screening of the Duchenne muscular dystrophyvia multiplex DNA apmlification. Nucleic Acids Res16: 11141
Fujimura FK, Northrup H, Beaudet AL, O'Brien WE (1990): Genotyping errors with the polymerase chain reaction. N Engl J Med322: 61
Fujishita S, Shibuya N, Niikawa N, Nagataki S (1991): Gene-deletion and carrier detections, and prenatal diagnosis of Duchenne muscular dystrophy by analysis of the dystrophin gene amplified by polymerase chain reaction. Jpn J Human Genet36: 317–324
Kitoh Y, Matsuo M, Nishio H, Takumi T, Nakajima T, Masumura T, Koga J, Nakamura H (1992): Amplification of the deletion rich exons of the dystrophin gene by polymerase chain reaction shows deletions in 36 of 90 Japanese families with Duchenne muscular dystrophy. Am J Med Genet42: 453–457
Koenig M, Hoffman EP, Bertelson CJ, Monaco AP, Feener C, Kunkel LM (1987): Complete cloning of the Duchenne muscular dystrophy (DMD) cDNA and gene in normal and affected individuals. Cell50: 509–517
Minami R (1991): Heterogeneity of gene deletions and clinical manifestations among DMD patients in Hokkaido. Annual report of research on the genetic, epidermiologic, clinical, and therapeutic aspects of muscular dystrophy supported by the National Center for Neurology and Psychiatry (NCNP) of the Ministry of Health and Welfare, Japan. 9–12.
Monaco AP, Bertelson CJ, Middlesworth W, Colletti CA, Aldridge J, Fishbeck KH, Bartlett R, Pericak-Vance MA, Roses AD, Kurkel LM (1985): Detection of deletions spanning the Duchenne muscular dystrophy locus using a tightly linked DNA segment. Nature316: 842–845
Moser H (1984): Duchenne muscular dystrophy: Pathogenic aspects and genetic prevention. Hum Genet66: 17–40
Multicenter Study Group (1992): Diagnosis of Duchenne and Becker muscular dystrophy by polymerase chain reaction. JAMA267: 2609–2615
Roberts RG, Cole CG, Hart KA, Bobrow M, Bentley DR (1989): Rapid carrier and prenatal diagnosis of Duchenne and Becker muscular dystrophy. Nucleic Acids Res17: 811
Sannomiya K (1991): DNA diagnosis of progressive muscular dystrophy by PCR technology. Annual report of research on the genetic, epidermiologic, clinical, and therapeutic aspects of muscular dystrophy supported by the National Center for Neurology and Psychiatry (NC-NP) of the Ministry of Health and Welfare, Japan. 65–69
Sugino S, Fujishita S, Kamimura N, Matsumoto T, Wapenaar MC, Deng HX, Shinuya N, Miike T, Niikawa N (1989): Molecular-genetic study of Duchenne and Becker muscular dystrophies: Deletion analyses of 45 Japanese patients and segregation analyses in their families with RF-LPs based on the data from normal Japanese females. Am J Med Genet34: 555–561
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Katayama, S., Takeshita, N., Yano, T. et al. Deletion detection for diagnosis of Duchenne muscular dystrophy in the Japanese population—Comparisom between the polymerase chain reaction and the Southern blot analysis. Jap J Human Genet 38, 177–184 (1993). https://doi.org/10.1007/BF01883708
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/BF01883708