MS-MLPA analysis for FMR1 gene: evaluation in a routine diagnostic setting
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- 作者：Valentina Gatta (1) (2)
Elena Gennaro (3)
Sara Franchi (1) (4)
Massimiliano Cecconi (3)
Ivana Antonucci (1)
Marco Tommasi (1)
Giandomenico Palka (4)
Domenico Coviello (3)
Liborio Stuppia (1) (2)
Marina Grasso (3) - 关键词：FMR1 ; Fragile X Syndrome ; Methylation ; MS ; MLPA
- 刊名：BMC Medical Genetics
- 出版年：2013
- 出版时间：December 2013
- 年：2013
- 卷：14
- 期：1
- 全文大小：389KB
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20. The pre-publication history for this paper can be accessed here: f="http://www.biomedcentral.com/1471-2350/14/79/prepub" class="a-plus-plus">http://www.biomedcentral.com/1471-2350/14/79/prepub - 作者单位：Valentina Gatta (1) (2)
Elena Gennaro (3)
Sara Franchi (1) (4)
Massimiliano Cecconi (3)
Ivana Antonucci (1)
Marco Tommasi (1)
Giandomenico Palka (4)
Domenico Coviello (3)
Liborio Stuppia (1) (2)
Marina Grasso (3)
1. Laboratory of Molecular Genetics, Department of Psychological, Humanities and Territorial Sciences, School of Medicine and Health Sciences, “G. d’Annunzio-University, via dei Vestini 31, Chieti, 66013, Italy
2. Aging Research Center, “G. d’Annunzio-Foundation, Chieti –Pescara, via Colle dell’Ara, Chieti, 66013, Italy
3. Laboratory of Genetics, Galliera Hospital, via A. Volta 8, Genoa, 16128, Italy
4. Department of Medical Sciences Oral and Biotechnologies, “G. d’Annunzio-University, via dei Vestini 31, Chieti, 66013, Italy - ISSN：1471-2350
文摘
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.