Combining approach with multiplex PCR and MLPA to detect deletion and duplication in DMD patients, carriers, and prenatal diagnosis / 中华医学遗传学杂志
Chinese Journal of Medical Genetics
;
(6): 318-322, 2009.
Article
in Chinese
| WPRIM
| ID: wpr-287399
ABSTRACT
<p><b>OBJECTIVE</b>Applying multiplex PCR and multiplex ligation-dependent probe amplification (MLPA) in a clinical setting to detect deletions and duplications in the Duchenne/Becker muscular dystrophy (DMD/BMD) gene not only for patients, but also for identification of possible carriers and prenatal diagnosis.</p><p><b>METHODS</b>Multiplex PCR was used first in patients clinically diagnosed with DMD/BMD to examine 26 exons for a large deletion in the two hot regions of the dystrophin gene. For patients without a deletion detected in the aforementioned regions, MLPA was used to further examine all 79 exons to determine whether a deletion in the remaining non-hot regions or any duplication was present. A similar approach was applied to suspected carriers. In requested prenatal diagnosis cases, specific PCR was used to detect deletions, while MLPA was applied to detect duplications.</p><p><b>RESULTS</b>Multiplex PCR was used to examine 26 exons within the two hot regions in the Dystrophin gene for 22 patients with DMD; 13 (13/22) had multi-exon deletions. For the 9 patients without deletions in the 26 exons, MLPA was used to examine 79 exons. 3 patients had duplications, 1 patient had a single deletion in exon 18, and no deletions or duplications could be detected in the remaining 5 patients. Of the 16 carriers, 2 out of the 3 that had family history had deletions, while the other 13 carriers were mothers of affected children who were sporadic patients without family history. Of them, 8 mothers were carriers for either deletions or duplications. For prenatal diagnosis, 9 fetuses were examined (one case was twins). Of them, 2 fetuses had familial deletions or duplications detected. These results were verified after induced abortion. In 7 fetuses, no deletions or duplications were detected and all developed into children.</p><p><b>CONCLUSION</b>Multiplex PCR can detect 92.86% of deletions and is useful for prenatal diagnosis of deletions because it is simple, reliable and inexpensive. It can be the first choice in DMD/BMD gene diagnosis. MLPA is important for detecting deletions in non-hot regions/exons and duplications in the DMD/BMD gene, as well as for carrier detection.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Prenatal Diagnosis
/
Polymerase Chain Reaction
/
Exons
/
Dystrophin
/
Sequence Deletion
/
Sequence Analysis, DNA
/
Gene Deletion
/
Clinical Laboratory Techniques
/
Muscular Dystrophy, Duchenne
/
Diagnosis
Type of study:
Diagnostic study
/
Prognostic study
Limits:
Adolescent
/
Adult
/
Child
/
Child, preschool
/
Female
/
Humans
/
Male
/
Pregnancy
Language:
Chinese
Journal:
Chinese Journal of Medical Genetics
Year:
2009
Type:
Article
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