Use of the Polymerase Chain Reaction and Complementary DNA Probes in theDetection of Duchenne Muscular Dystrophy

摘要

Duchenne Muscular Dystrophy (DMD) is a lethal X-linked myopathy occurring in 1 in3000 male births. The gene, which spans over 2.1 kilobases, has been identified and produces a protein designated dystrophin which is an integral component of the cytoskeleton of muscle membrane. In 50 to 60 % of DMD males, the gene defect is a deletion which disrupts the reading frame and results in significantly reduced production or abnormal structure of dystrophin. The objective of the first phase of this research was to identify deletions in the dystrophin gene in DMD males using a polymerase chain reaction (PCR) procedure. In the PCR, specific oligonucleotides are used to selectively amplify target sites in the gene up to a million fold. In this protocol 9 unique oligonucleotide primers were used in a multiplex PCR to simultaneously amplify sites within the gene known to be deletion prone.