Identification of female carriers for Duchenne and Becker muscular dystrophies using a FISH-based approach

Ligon, Azra H.; Kashork, Catherine D.; Richards, C. Sue; Shaffer, Lisa G.

doi:10.1038/sj.ejhg.5200450

Cited by 24 publications

(11 citation statements)

References 15 publications

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“…The above described PCR approach is not useful in detecting female carriers in whom deletions are masked by the amplification of the normal X chromosome. Thus, identification of female carriers must be carried out by different strategies, such as linkage analysis with intragenic polymorphic markers (Darras et al 1987;Clemens et al 1991), quantitative analysis of gene dosage (Prior et al 1990;Abbs and Bobrow 1993) and fluorescence in situ hybridisation (FISH) analysis (Voskova-Goldman et al 1997;Ligon et al 2000). However, linkage analysis cannot be used in families with a single affected male or performed if DNA of the affected members of the family is not available.…”

Section: Introductionmentioning

confidence: 97%

Identification of deletions and duplications of the DMD gene in affected males and carrier females by multiple ligation probe amplification (MLPA)

et al. 2005

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Duchenne muscular dystrophy (DMD) and Becker muscular dystrophy (BMD) are caused in the majority of cases by deletions of the DMD gene and are readily detectable in affected males by multiplex polymerase chain reaction (PCR). However, different approaches must be used for the identification of female carriers, in which deletions are not detectable by PCR, because of the presence of a normal X chromosome. In this study, we used the multiple ligation probe amplification (MLPA) tool for the identification of female carriers of DMD deletions or duplications in 12 families with a single affected male, 10 of which were previously diagnosed as carriers of a DMD rearrangement, and the remaining two as having an unknown disease-causing mutation. In all the investigated affected males, MLPA analysis confirmed the presence of a DMD rearrangement, and in six of them allowed the refinement of the breakpoints. In 12 female relatives of the affected patients, MLPA analysis showed a DMD deletion or duplication, confirming their carrier status. Two of these were the mother and the sister of a patient whose disease-causing mutation was not known. MLPA analysis thus proved to be an useful tool for the analysis of both affected males and females carriers of DMD rearrangements in cases in which the disease-causing mutation in the affected male was not known, providing useful information for the genetic counselling of the family.

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Section: Introductionmentioning

confidence: 97%

Identification of deletions and duplications of the DMD gene in affected males and carrier females by multiple ligation probe amplification (MLPA)

et al. 2005

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“…Thus, while confirming the results of other groups, FISH using DMD exon-specific probes represents an effective, highly accurate, direct, qualitative approach for establishing DMD carrier status of females (Ligon et al, 2000;Hermanova et al, 2002). Even when there is no cure for DMD patients, providing an accurate diagnosis for female carriers allows bringing the family to a precise genetic counseling.…”

mentioning

confidence: 59%

“…These molecular tools are very useful for detecting DMD gene deletions in males, but for females the detection of deletions can be problematic since women possess two X chromosomes (Ligon et al, 2000). More recently, FISH detecting chromosomal deletions has been introduced to identify specific DMD exon deletions (Voskova-Goldman et al, 1997;Ligon et al, 2000;Hermanova et al, 2002).…”

Section: Discussionmentioning

confidence: 95%

“…However, some groups have used the fluorescence in situ hybridization (FISH) assay to identify DMD female carriers since it provides an alternative approach to current molecular methods for carrier detection (Voskova-Goldman et al, 1997;Ligon et al, 2000;Hermanova et al, 2002). Importantly, this method is very useful to detect deletions since it is directed at exon-specific hot spots of the dystrophin gene, which are responsible for the majority of deletions.…”

Section: Introductionmentioning

confidence: 97%

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Identification of Duchenne Muscular Dystrophy Female Carriers by Fluorescence In Situ Hybridization and RT-PCR

et al. 2008

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Duchenne muscular dystrophy (DMD) is an X-linked recessive neuromuscular disorder caused by mutations in the dystrophin DMD gene located at Xp21.1 region. Up to 65% of the patients present dystrophin gene deletions. Mothers of DMD patients have a two-thirds chance of carrying a dystrophin mutation. The female carrier will transmit the disease gene to half of her sons and half of her daughters. As the recurrence risk for the disease is extremely high, it is very important to detect carrier status among female relatives of the patients to bring an adequate genetic counseling. In this work, results from two methods to identify female carriers are presented. One method is a multicolor fluorescence in situ hybridization (FISH) assay, and the other is reverse transcriptase-polymerase chain reaction (RT-PCR). We showed that FISH is an efficient, sensitive method that brings confident results to detect DMD female carriers as compared to RT-PCR.

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“…The above described PCR approach is unable to detect either duplications or female carriers in whom deletions are masked by the amplification of the normal X chromosome. Thus, identification of female carriers requires different strategies, such as segregation analysis with intragenic polymorphic markers (Darras et al, 1987;Clemens et al, 1991), quantitative analysis of gene dosage (Prior et al, 1990;Abbs and Bobrow, 1993) and fluorescence in situ hybridization (FISH) analysis (Voskova-Goldman et al, 1997;Ligon et al, 2000). However, segregation analysis cannot be used in families with a single affected male or performed if DNA of the affected members of the family is not available.…”

Section: Introductionmentioning

confidence: 99%

Detection of deletions and duplications in the Duchenne muscular dystrophy gene by the molecular method MLPA in the first Argentine affected families

Marzese

Mampel²,

Gómez³

et al. 2008

Genet. Mol. Res.

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AbstRACt. Deletions/duplications in the Duchenne muscular dystrophy (DMD) gene account for 60 to 70% of all alterations. A new technique, multiplex ligation-dependent probe amplification (MLPA), has been described that allows the detection of large genetic rearrangements by simultaneous amplification of up to 45 target sequences. The present article is based on the diagnosis of the first Argentine affected families by the application of MLPA. DNA samples from patients with and without a previous diagnosis were included. MLPA assays were performed according to manufacturer recommendations. Polymerase chain reaction and direct sequencing were performed when a single-exon deletion was detected. Results were analyzed using the Gene Marker v1.6 and Sequencing Analysis v5.2 software. In the samples with a previous diagnosis (as identified by short tandem repeat-polymerase chain reaction analysis), MLPA confirmed in some samples the same deletion and detected in others a larger deleted fragment. This enabled the prediction of the expected male phenotype. One deletion and one duplication were detected in patients without previous diagnosis. In this study, we investigated the applicability of MLPA in our country. Our results showed a 100% confirmation of the deleted fragments detected by short tandem repeat segregation analysis. Moreover, in some cases, the MLPA assay was able to refine the breakpoints involved. In addition, MLPA identified deletions/duplications in samples without previous diagnosis. In comparison to the available diagnosis strategies in Argentina, MLPA is less time-consuming, and spans the complete coding region of DMD. The application of MLPA will improve the genetic diagnosis of DMD/Becker muscular dystrophy in our country.

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Identification of female carriers for Duchenne and Becker muscular dystrophies using a FISH-based approach

Cited by 24 publications

References 15 publications

Identification of deletions and duplications of the DMD gene in affected males and carrier females by multiple ligation probe amplification (MLPA)

Identification of deletions and duplications of the DMD gene in affected males and carrier females by multiple ligation probe amplification (MLPA)

Identification of Duchenne Muscular Dystrophy Female Carriers by Fluorescence In Situ Hybridization and RT-PCR

Detection of deletions and duplications in the Duchenne muscular dystrophy gene by the molecular method MLPA in the first Argentine affected families

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