Protein truncation test (PTT) to rapidly screen the DMD gene for translation terminating mutations

Roest, Pauline A.M.; Roberts, Roland G.; Tuijn, A.C. van der; Heikoop, Judith C.; Ommen, Gert‐Jan B. van; Dunnen, Johan T. den

doi:10.1016/0960-8966(93)90083-v

Cited by 57 publications

(24 citation statements)

References 2 publications

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“…As its performance clearly outperforms the Beggs and Chamberlain multiplex-PCR test 6,7 it should be considered as the method of choice for a first DNA analysis of DMD/BMD patients. After a negative MLPA-test, the gene can be scanned for point mutations using RNA 21,22 or DNA-based tests. 23 -25 …”

Section: Discussionmentioning

confidence: 99%

Deletion and duplication screening in the DMD gene using MLPA

et al. 2005

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We have designed a multiplex ligation-dependent probe amplification (MLPA) assay to simultaneously screen all 79 DMD gene exons for deletions and duplications in Duchenne and Becker muscular dystrophy (DMD/BMD) patients. We validated the assay by screening 123 unrelated patients from Serbia and Montenegro already screened using multiplex PCR. MLPA screening confirmed the presence of all previously detected deletions. In addition, we detected seven new deletions, nine duplications, one point mutation, and we were able to precisely determine the extent of all rearrangements. To facilitate MLPAbased screening in laboratories lacking specific equipment, we designed the assay such that it can also be performed using agarose gel analysis and ethidium bromide staining. The MLPA assay as described provides a simple and cheap method for deletion and duplication screening in DMD/BMD patients. The assay outperforms the Beggs and Chamberlain multiplex-PCR test, and should be considered as the method of choice for an initial DNA analysis of DMD/BMD patients.

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Section: Discussionmentioning

confidence: 99%

Deletion and duplication screening in the DMD gene using MLPA

et al. 2005

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“…Numerous methods have been applied to scan the DMD gene for nucleotide changes, including: SSCP [11], dHPLC [12], FM-CSCE [13], PTT [14], HRM [15]. These pre-screens aim to offer a lower cost alternative to sequencing all the 79 DMD exons, however, since the cost of sequencing has reduced dramatically over recent years it may be more appropriate to sequence the full gene now [16].…”

Section: Testing For Other Mutation Typesmentioning

confidence: 99%

“…A meeting of 29 senior scientists from Europe, the USA, India and Australia, was held in Naarden, The Netherlands on November [14][15][16]2008, to establish consensus Best Practice Guidelines for molecular diagnosis of Duchenne and Becker muscular dystrophy (DMD/BMD). New therapeutic trials for DMD demand accurate diagnosis of the disorder, especially where the therapy is targeted towards specific mutations.…”

Section: Introductionmentioning

confidence: 99%

Best Practice Guidelines on molecular diagnostics in Duchenne/Becker muscular dystrophies

Abbs

Tuffery‐Giraud

Bakker

et al. 2010

Neuromuscular Disorders

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“…Hence, there is extensive reliance on rapid screening techniques to quickly detect the subset of samples with apparent changes and that therefore merit sequencing. Many techniques have been developed for the detection of DNA sequence variations including the protein truncation test (PTT), 1 denaturing gradient gel electrophoresis (DGGE), 2 chemical cleavage of mismatches, 3 single-strand conformation polymorphism (SSCP), 4 2-dimensional gene scanning, 5 denaturing high-performance liquid chromatography (D-HPLC), 6 and conformation-sensitive gel electrophoresis (CSGE). 7 A detailed evaluation of the advantages and disadvantages of the current techniques indicates that some rapid screening techniques are too expensive (DGGE) or are not amenable to high throughput (DGGE and PTT).…”

mentioning

confidence: 99%

Conformation-Sensitive Gel Electrophoresis as an Ideal High-Throughput Strategy for Accurate Detection of Sequence Variations in DNA: Screening hTomm and hTimm Genes

Blesa¹,

Prieto-Ruiz²,

Hernández-Yago³

2004

SLAS Discovery

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Conformation-sensitive gel electrophoresis is a heteroduplex-based method that is particularly well suited to high-throughput analyses. Its simplicity makes it amenable to various adaptations and modifications to enhance its applicability to genome-wide mutation scans. Technical aspects that markedly improve the conformation-sensitive gel electrophoresis performance by combining high throughput and high resolution of the bands facilitating the interpretation of the results are described here. The authors report some of the results they have obtained in the screening of the exon 1 of human Timm8A gene as an example of the suitability of the conformation-sensitive gel electrophoresis–based protocol that has been adapted to optimize its throughput, speed, and simplicity in the recognition of both heterozygous and homozygous DNA mutations. The higher throughput is achieved by using 12 batches per gel. The length of the gel is sufficient for an adequate well-to-read distance for each batch that allows a clear distinction and resolution of the conformation-sensitive gel electrophoresis bands. Standardization of the procedure using multichannel pipettes reduces the preparation time of the 96-well PCRs to 10 min and also accelerates the gel loading. The resulting bands give high-quality images, allowing easy detection of known as well as novel mutations.

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Protein truncation test (PTT) to rapidly screen the DMD gene for translation terminating mutations

Cited by 57 publications

References 2 publications

Deletion and duplication screening in the DMD gene using MLPA

Deletion and duplication screening in the DMD gene using MLPA

Best Practice Guidelines on molecular diagnostics in Duchenne/Becker muscular dystrophies

Conformation-Sensitive Gel Electrophoresis as an Ideal High-Throughput Strategy for Accurate Detection of Sequence Variations in DNA: Screening hTomm and hTimm Genes

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