title: Automated high multiplex qPCR platform for simultaneous detection and quantification of multiple nucleic acid targets.
abstract: Quantitative PCR (qPCR) using real-time detection of amplification is limited to a small number of targets within a single reaction. The ICEPlex system, using our scalable target analysis routine (STAR) technology, was developed to provide an automated, high multiplexing PCR solution. ICEPlex combines PCR thermal cycling with dynamic, sequential amplicon separation by capillary electrophoresis and two-color quantitative detection in a single integrated system. In contrast to probe-based qPCR, ICEPlex directly measures amplicon accumulation through incorporation of labeled primers. Three orders of magnitude of optical detection range and at least 7 logs of detectable target concentration range are demonstrated. The system can separate more than 50 amplicons per color channel, ranging from 100 to 500 bases, providing broad multiplexing capabilities for a wide spectrum of nucleic acid amplification applications. ICEPlex can be used for analysis of viral DNA or RNA targets, detection of genetic variants, and for reverse-transcriptase PCR gene expression panels.
authors.affiliation: PrimeraDx, Mansfield, MA, USA.
authors.forename: Louis
authors.initials: L
authors.lastname: Hlousek
authors.forename: Sergey
authors.initials: S
authors.lastname: Voronov
authors.forename: Vesselin
authors.initials: V
authors.lastname: Diankov
authors.forename: Amy B
authors.initials: AB
authors.lastname: Leblang
authors.forename: Patrick J
authors.initials: PJ
authors.lastname: Wells
authors.forename: Donna M
authors.initials: DM
authors.lastname: Ford
authors.forename: Jork
authors.initials: J
authors.lastname: Nolling
authors.forename: Kyle W
authors.initials: KW
authors.lastname: Hart
authors.forename: Patricio A
authors.initials: PA
authors.lastname: Espinoza
authors.forename: Michael R
authors.initials: MR
authors.lastname: Bristol
authors.forename: Gregory J
authors.initials: GJ
authors.lastname: Tsongalis
authors.forename: Belinda
authors.initials: B
authors.lastname: Yen-Lieberman
authors.forename: Vladimir I
authors.initials: VI
authors.lastname: Slepnev
authors.forename: Lilly I
authors.initials: LI
authors.lastname: Kong
authors.forename: Ming-Chou
authors.initials: MC
authors.lastname: Lee
mesh_terms: D003587:Cytomegalovirus / Q000235:genetics; D004279:DNA, Viral / Q000097:blood; D019075:Electrophoresis, Capillary / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry; D004854:Herpesvirus 4, Human / Q000235:genetics; D006801:Humans; D016014:Linear Models; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D009696:Nucleic Acids / Q000032:analysis* / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: D004279:DNA, Viral; D005456:Fluorescent Dyes; D009696:Nucleic Acids