title: Three-color crystal digital PCR.
abstract: Digital PCR is an exciting new field for molecular analysis, allowing unprecedented precision in the quantification of nucleic acids, as well as the fine discrimination of rare molecular events in complex samples. We here present a novel technology for digital PCR, Crystal Digital PCR™, which relies on the use of a single chip to partition samples into 2D droplet arrays, which are then subjected to thermal cycling and finally read using a three-color fluorescence scanning device. This novel technology thus allows three-color multiplexing, which entails a different approach to data analysis. In the present publication, we present this innovative workflow, which is both fast and user-friendly, and discuss associated data analysis issue, such as fluorescence spillover compensation and data representation. Lastly, we also present proof-of-concept of this three-color detection system, using a quadriplex assay for the detection of EGFR mutations L858R, L861Q and T790M.
authors.affiliation: Stilla Technologies, 1 Mail du Professeur Georges Mathé, 94800 Villejuif, France.
authors.forename: J
authors.initials: J
authors.lastname: Madic
authors.affiliation: Stilla Technologies, 1 Mail du Professeur Georges Mathé, 94800 Villejuif, France.
authors.forename: A
authors.initials: A
authors.lastname: Zocevic
authors.affiliation: Ecole Polytechnique, Route de Saclay, 91128 Palaiseau, France.
authors.forename: V
authors.initials: V
authors.lastname: Senlis
authors.affiliation: Stilla Technologies, 1 Mail du Professeur Georges Mathé, 94800 Villejuif, France.
authors.forename: E
authors.initials: E
authors.lastname: Fradet
authors.affiliation: Stilla Technologies, 1 Mail du Professeur Georges Mathé, 94800 Villejuif, France.
authors.forename: B
authors.initials: B
authors.lastname: Andre
authors.affiliation: Stilla Technologies, 1 Mail du Professeur Georges Mathé, 94800 Villejuif, France.
authors.forename: S
authors.initials: S
authors.lastname: Muller
authors.affiliation: Stilla Technologies, 1 Mail du Professeur Georges Mathé, 94800 Villejuif, France.
authors.forename: R
authors.initials: R
authors.lastname: Dangla
authors.affiliation: Stilla Technologies, 1 Mail du Professeur Georges Mathé, 94800 Villejuif, France.
authors.forename: M E
authors.initials: ME
authors.lastname: Droniou
keywords: Crystal Digital PCR; EGFR; Multicolor PCR; Multiplex digital PCR; NSCLC; Naica