Evaluation of Multiplex PCR Assay Using Dual Priming Oligonucleotide System for Detection Mutation in the Duchenne Muscular Dystrophy Gene.
10.3343/kjlm.2008.28.5.386
- Author:
Younhee PARK
1
;
Juwon KIM
;
Jong Rak CHOI
;
Jaewoo SONG
;
Jong Shin CHUNG
;
Kyung A LEE
Author Information
1. Department of Laboratory Medicine, Yonsei University College of Medicine, Seoul, Korea. KAL1119@yuhs.ac
- Publication Type:Original Article ; Comparative Study ; English Abstract ; Evaluation Studies
- Keywords:
DMD gene;
Deletion;
Multiplex PCR;
Dual priming oligonucleotide PCR;
Multiplex ligation-dependent probe amplification
- MeSH:
*DNA Mutational Analysis;
DNA Primers;
Dystrophin/*genetics;
Female;
Gene Deletion;
Genetic Screening;
Humans;
Male;
Muscular Dystrophy, Duchenne/*diagnosis/genetics;
Nucleic Acid Amplification Techniques;
Oligonucleotide Probes;
Polymerase Chain Reaction/*methods;
Reagent Kits, Diagnostic;
Reproducibility of Results
- From:The Korean Journal of Laboratory Medicine
2008;28(5):386-391
- CountryRepublic of Korea
- Language:Korean
-
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.
