[Evaluation of multiplex PCR assay using dual priming oligonucleotide system for detection mutation in the Duchenne muscular dystrophy gene].
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.
Park
Younhee
Y
Department of Laboratory Medicine, Yonsei University College of Medicine, Seoul, Korea.
Kim
Juwon
J
Choi
Jong Rak
JR
Song
Jaewoo
J
Chung
Jong Shin
JS
Lee
Kyung A
KA
D004252:DNA Mutational Analysis; D017931:DNA Primers; D016189:Dystrophin / Q000235:genetics*; D005260:Female; D017353:Gene Deletion; D005820:Genetic Testing; D006801:Humans; D008297:Male; D020388:Muscular Dystrophy, Duchenne / Q000175:diagnosis* / Q000235:genetics; D021141:Nucleic Acid Amplification Techniques; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction / Q000379:methods*; D011933:Reagent Kits, Diagnostic; D015203:Reproducibility of Results
C484258:DMD protein, human; D017931:DNA Primers; D016189:Dystrophin; D015345:Oligonucleotide Probes; D011933:Reagent Kits, Diagnostic