µLAS: Sizing of expanded trinucleotide repeats with femtomolar sensitivity in less than 5 minutes.

Details

Ressource 1Download: s41598-018-36632-5.pdf (2282.61 [Ko])
State: Public
Version: Final published version
License: Not specified
Serval ID
serval:BIB_B0AED8A58FBC
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
µLAS: Sizing of expanded trinucleotide repeats with femtomolar sensitivity in less than 5 minutes.
Journal
Scientific reports
Author(s)
Malbec R., Chami B., Aeschbach L., Ruiz Buendía G.A., Socol M., Joseph P., Leïchlé T., Trofimenko E., Bancaud A., Dion V.
ISSN
2045-2322 (Electronic)
ISSN-L
2045-2322
Publication state
Published
Issued date
10/01/2019
Peer-reviewed
Oui
Volume
9
Number
1
Pages
23
Language
english
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't
Publication Status: epublish
Abstract
We present µLAS, a lab-on-chip system that concentrates, separates, and detects DNA fragments in a single module. µLAS speeds up DNA size analysis in minutes using femtomolar amounts of amplified DNA. Here we tested the relevance of µLAS for sizing expanded trinucleotide repeats, which cause over 20 different neurological and neuromuscular disorders. Because the length of trinucleotide repeats correlates with the severity of the diseases, it is crucial to be able to size repeat tract length accurately and efficiently. Expanded trinucleotide repeats are however genetically unstable and difficult to amplify. Thus, the amount of amplified material to work with is often limited, making its analysis labor-intensive. We report the detection of heterogeneous allele lengths in 8 samples from myotonic dystrophy type 1 and Huntington disease patients with up to 750 CAG/CTG repeats in five minutes or less. The high sensitivity of the method allowed us to minimize the number of amplification cycles and thus reduce amplification artefacts without compromising the detection of the expanded allele. These results suggest that µLAS can speed up routine molecular biology applications of repetitive sequences and may improve the molecular diagnostic of expanded repeat disorders.
Keywords
Diagnostic Tests, Routine/methods, Humans, Lab-On-A-Chip Devices, Nervous System Diseases/diagnosis, Nervous System Diseases/pathology, Neuromuscular Diseases/diagnosis, Neuromuscular Diseases/pathology, Sensitivity and Specificity, Trinucleotide Repeat Expansion
Pubmed
Web of science
Open Access
Yes
Create date
28/01/2019 9:11
Last modification date
30/01/2020 6:20
Usage data