| Title: |
A Simple, Universal, and Cost-Efficient Digital PCR Method for the Targeted Analysis of Copy Number Variations |
| Authors: |
Cassinari, Kévin; Quenez, Olivier; Joly-Hélas, Géraldine; Beaussire, Ludivine; Le Meur, Nathalie; Castelain, Mathieu; Goldenberg, Alice; Guerrot, Anne-Marie; Brehin, Anne-Claire; Deleuze, Jean-François; Boland, Anne; Rovelet-Lecrux, Anne; Campion, Dominique; Saugier-Veber, Pascale; Gruchy, Nicolas; Frebourg, Thierry; Nicolas, Gaël; Sarafan-Vasseur, Nasrin; Chambon, Pascal |
| Source: |
Clinical Chemistry ; volume 65, issue 9, page 1153-1160 ; ISSN 0009-9147 1530-8561 |
| Publisher Information: |
Oxford University Press (OUP) |
| Publication Year: |
2019 |
| Description: |
BACKGROUND Rare copy number variations (CNVs) are a major cause of genetic diseases. Simple targeted methods are required for their confirmation and segregation analysis. We developed a simple and universal CNV assay based on digital PCR (dPCR) and universal locked nucleic acid (LNA) hydrolysis probes. METHODS We analyzed the mapping of the 90 LNA hydrolysis probes from the Roche Universal ProbeLibrary (UPL). For each CNV, selection of the optimal primers and LNA probe was almost automated; probes were reused across assays and each dPCR assay included the CNV amplicon and a reference amplicon. We assessed the assay performance on 93 small and large CNVs and performed a comparative cost-efficiency analysis. RESULTS UPL-LNA probes presented nearly 20000000 occurrences on the human genome and were homogeneously distributed with a mean interval of 156 bp. The assay accurately detected all the 93 CNVs, except one ( |
| Document Type: |
article in journal/newspaper |
| Language: |
English |
| DOI: |
10.1373/clinchem.2019.304246 |
| Availability: |
https://doi.org/10.1373/clinchem.2019.304246; http://academic.oup.com/clinchem/article-pdf/65/9/1153/33806472/clinchem1153.pdf |
| Rights: |
https://academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model |
| Accession Number: |
edsbas.A920D297 |
| Database: |
BASE |