JMD GMP oligos for in vitro Diagnostics
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Originally published online as doi:10.2353/jmoldx.2009.080094 on February 5, 2009

Published online before print February 5, 2009
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Journal of Molecular Diagnostics 2009, Vol. 11, No. 2
Copyright © 2009 American Society for Investigative Pathology & Association for Molecular Pathology
DOI: 10.2353/jmoldx.2009.080094

Award Lecture

LightCycler Technology in Molecular Diagnostics

Elaine Lyon and Carl T. Wittwer

From the Department of Pathology, University of Utah, and the ARUP Institute for Experimental and Clinical Pathology, Salt Lake City, Utah

Abstract

LightCycler technology combines rapid-cycle polymerase chain reaction with real-time fluorescent monitoring and melting curve analysis. Since its introduction in 1997, it is now used in many areas of molecular pathology, including oncology (solid tumors and hematopathology), inherited disease, and infectious disease. By monitoring product accumulation during rapid amplification, quantitative polymerase chain reaction in a closed-tube system is possible in 15 to 30 minutes. Furthermore, melting curve analysis of probes and/or amplicons provides genotyping and even haplotyping. Novel mutations are identified by unexpected melting temperature or curve shape changes. Melting probe designs include adjacent hybridization probes, single labeled probes, unlabeled probes, and snapback primers. High-resolution melting allows mutation scanning by detecting all heterozygous changes. This review describes the major advances throughout the last 15 years regarding LightCycler technology and its application in clinical laboratories.




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