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

Rémi Malbec (1) , Bayan Chami (2) , Lorène Aeschbach (3) , Gustavo Ruiz Buendía (3) , Marius Socol (2) , Pierre Joseph (2) , Thierry Leichle (4) , Evgeniya Trofimenko (3) , Aurélien Bancaud (2) , Vincent Dion (3)

1 LAAS-MICA - Équipe MICrosystèmes d'Analyse
2 LAAS-MILE - Équipe Micro-Nanofluidique pour les sciences de la vie et de l’environnement
3 UNIL - Université de Lausanne = University of Lausanne
4 LAAS-MEMS - Équipe Microsystèmes électromécaniques

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.

Domaines: Micro et nanotechnologies/Microélectronique; Génomique, Transcriptomique et Protéomique [q-bio.GN]

Rémi Malbec, Bayan Chami, Lorène Aeschbach, Gustavo Ruiz Buendía, Marius Socol, et al.. µLAS: Sizing of expanded trinucleotide repeats with femtomolar sensitivity in less than 5 minutes. Scientific Reports, 2019, 9, pp.23. ⟨10.1038/s41598-018-36632-5⟩. ⟨hal-01997319⟩