Mutation detection using fluorescent hybridization probes and melting curve analysis

Abstract

The LightCycler^® is a real-time PCR instrument that combines a thermocycler and a micro-volume fluorimeter. LightCycler technology is gaining popularity due to its ability to detect mutations quickly and accurately. Multiple base alterations are discriminated using hybridization probes and fluorescent melting curves. This review focuses on mutation detection and base discrimination by fluorescent hybridization probes. Assay designs for single base mutation detection and complex multiplex reactions are discussed. Types of mutations detected and reported applications are reviewed. Guidelines using melting curve analysis for the clinical laboratory are presented.

• fluorescent monitoring
• hybridization probes
• LightCycler
• melting curves
• mutation detection