Automated high multiplex qPCR platform for simultaneous detection and quantification of multiple nucleic acid targets

BioTechniques
Louis HlousekMing-Chou Lee

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.

References

Aug 15, 1991·Proceedings of the National Academy of Sciences of the United States of America·P M HollandD H Gelfand
Jun 25, 1983·Nucleic Acids Research·M D Jones, B E Griffin
Mar 9, 2002·Nucleic Acids Research·Ian M MackayAndreas Nitsche
Apr 4, 2002·Annals of Clinical Biochemistry·Larry J Kricka
Oct 21, 2004·Journal of Clinical Virology : the Official Publication of the Pan American Society for Clinical Virology·Guy Vernet
Nov 4, 2008·Journal of Clinical Virology : the Official Publication of the Pan American Society for Clinical Virology·R N GunsonW F Carman
Feb 21, 2009·The New England Journal of Medicine·Christos Sotiriou, Lajos Pusztai

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Citations

Sep 10, 2015·Clinical Chemistry and Laboratory Medicine : CCLM·Dong-Sheng Lian, Shu-Jin Zhao

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Methods Mentioned

BETA
PCR
electrophoresis
ICE
chip
Assay
PCRs

Software Mentioned

ICE Plex
ViraQuant
STAR

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