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Clinical Chemistry 36: 2113-2117, 1990;
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Clinical Chemistry, Vol 36, 2113-2117, Copyright © 1990 by American Association for Clinical Chemistry

Determination of carrier status in Duchenne and Becker muscular dystrophies by quantitative polymerase chain reaction and allele- specific oligonucleotides

TW Prior, AC Papp, PJ Snyder, WE Highsmith Jr, KJ Friedman, TR Perry, LM Silverman and JR Mendell
Department of Pathology, Ohio State University, Columbus 43210.

Detection of carriers of Duchenne muscular dystrophy (DMD) and Becker muscular dystrophy (BMD), in the deletion cases, involves calculating gene dosage from Southern blots. We show that the analysis of dosage can also be made from the polymerase chain reaction (PCR) with use of allele-specific oligonucleotides (ASOs). The deletion-prone exons are amplified, transferred to a membrane, and hybridized with ASOs complementary to the exons; the autoradiographic bands are then quantified with a densitometer. After determining the quantitative conditions of the amplification reaction, we were able to identify deletions in a DMD/BMD carrier female. The determination of carrier status via PCR removes several of the technical limitations of Southern analysis and is also cost- and labor-effective.


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K. J FLINTOFF, E. SHERIDAN, G. TURNER, C. E CHU, and G. R TAYLOR
Submicroscopic deletions of the APC gene: a frequent cause of familial adenomatous polyposis that may be overlooked by conventional mutation scanning
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T. W. Prior
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P. Fortina, J. Cheng, M. A. Shoffner, S. Surrey, W. M. Hitchcock, L. J. Kricka, and P. Wilding
Diagnosis of Duchenne/Becker muscular dystrophy and quantitative identification of carrier status by use of entangled solution capillary electrophoresis
Clin. Chem., May 1, 1997; 43(5): 745 - 751.
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