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Open Access 01-12-2013 | Research article

MS-MLPA analysis for FMR1 gene: evaluation in a routine diagnostic setting

Authors: Valentina Gatta, Elena Gennaro, Sara Franchi, Massimiliano Cecconi, Ivana Antonucci, Marco Tommasi, Giandomenico Palka, Domenico Coviello, Liborio Stuppia, Marina Grasso

Published in: BMC Medical Genetics | Issue 1/2013

Abstract

Background

Fragile X Syndrome (FXS), the most common cause of familiar mental retardation, is associated in over 99% of cases to an expansion over 200 repeats of a CGG sequence in the 5’ UTR of the FMR1 gene (Xq27.3), leading to the hypermethylation of the promoter. Molecular diagnosis of FXS have been so far based on the use of the Southern Blot (SB) analysis, a low throughput and time consuming technique. In order to update the diagnostic approach for FXS, we evaluated the usefulness of the Methylation-Specific Multiplex-Ligation-dependent Probe Amplification assay (MS-MLPA).

Methods

The study was carried out by retrospectively analysing 44 male patients, 10 Chorionic Villus Sampling (CVS) samples and 10 females previously analyzed by SB. In addition, a prospective study on 98 male subjects, 20 females and 1 CVS sample was carried out for assessing the feasibility and the impact of MS-MLPA in a routine lab work.

Result

Results provided by both the retrospective and the prospective parts of this study strongly demonstrate the robustness and reproducibility of the MS-MLPA assay, able to correctly detect the methylation status in all normal and full mutation male samples analyzed, including CVS male samples. On the other hand, MS-MLPA analysis on females samples produced unreliable results.

Conclusion

Based on our results, we suggest the necessity of a separate workflow for male and female patients with suspected FXS in the routine diagnostic setting. MS-MLPA, in combination with CGG repeat sizing using a single-tube primed FMR1 PCR, represents a reliable diagnostic protocol in the molecular diagnosis of FXS male patients.

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The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2350/14/79/prepub

Title: MS-MLPA analysis for FMR1 gene: evaluation in a routine diagnostic setting
Authors: Valentina Gatta
Elena Gennaro
Sara Franchi
Massimiliano Cecconi
Ivana Antonucci
Marco Tommasi
Giandomenico Palka
Domenico Coviello
Liborio Stuppia
Marina Grasso
Publication date: 01-12-2013
Publisher: BioMed Central
Published in: BMC Medical Genetics / Issue 1/2013
Electronic ISSN: 1471-2350
DOI: https://doi.org/10.1186/1471-2350-14-79

Keynote webinar | Spotlight on medication adherence

Springer Medicine

MS-MLPA analysis for FMR1 gene: evaluation in a routine diagnostic setting

Abstract

Background

Methods

Result

Conclusion

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Background

Methods

Result

Conclusion

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