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Abstract
Background
Exon deletions of Duchenne muscular dystrophy (DMD) gene account for most of the alterations found in DMD and Becker muscular dystrophy (BMD). This study was to evaluate the usefulness of dual priming oligonucleotide multiplex PCR (DPO PCR) in detection of exon deletions of DMD gene.
Methods
Thirty-seven DMD or BMD patients who had known exon deletions detected by conventional multiplex PCR (conventional PCR) and nine control subjects were enrolled in this study. When a discrepancy was shown between the results of conventional PCR and DPO PCR, the multiplex ligation-dependent probe amplification (MLPA) technique was performed as a confirmation test.
Results
The same deletions previously identified by conventional PCR in 32 out of 37 subjects were also detected by DPO PCR. For the five subjects (13.5%) showing discrepant results between the conventional PCR and DPO PCR, MLPA was performed and its results were found to correlate better with those of DPO PCR. The discrepancies were due to false positive or false negative results of the conventional PCR.
Conclusions
DPO PCR shows a high agreement of results with the conventional PCR and is considered an adequate method to be used as a primary genetic test for the diagnosis of DMD. Because of an improved accuracy, especially for determining the boundaries of DMD gene deletions, DPO PCR can be very useful as a supplement to the conventional PCR.
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Fig. 1.
Multiplex ligation-dependent probe amplification (MLPA), conventional-multiplex PCR, and dual priming oligonucleotide (DPO) multiplex PCR of case 34. (A) Deletion of exons 48 and 49 of the DMD gene (arrows) in MLPA. (B) Deletion of exon 49 in lane 2 and presence of exon 48 band in lane 1 of conventional-multiplex PCR. (C) Deletion of exon 48 in group 3 DPO multiplex PCR. M, the 100-bp-size marker.
Table 1.
Concordant results between conventional multiplex PCR and DPO multiplex PCR
Table 2.
Results of MLPA performed on the five cases that produced discrepant results between conventional-multiplex PCR and DPO multiplex PCR
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