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  • Volume 52, Issue 10
  • Rapid detection of the factor V Leiden (1691 G > A) and haemochromatosis (845 G > A) mutation by fluorescence resonance energy transfer (FRET) and real time PCR.

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Rapid detection of the factor V Leiden (1691 G > A) and haemochromatosis (845 G > A) mutation by fluorescence resonance energy transfer (FRET) and real time PCR.
  1. S H Neoh,
  2. M J Brisco,
  3. F A Firgaira,
  4. K J Trainor,
  5. D R Turner,
  6. A A Morley
  1. Department of Haematology and Genetic Pathology, Flinders University of South Australia, Flinders Medical Centre, Australia.

    Abstract

    A rapid method based on fluorescence resonance energy transfer (FRET) and real time polymerase chain reaction (PCR) was used to identify the haemochromatosis genotype in 112 individuals and the factor V genotype in 134 individuals. The results were compared with conventional methods based on restriction enzyme digestion of PCR products. The two methods agreed in 244 of the 246 individuals; for the other two individuals, sequencing showed that they had been incorrectly genotyped by the standard method but correctly genotyped by FRET. The simplicity, speed, and accuracy of real time PCR analysis using FRET probes make it the method of choice in the clinical laboratory for genotyping the haemochromatosis and factor V genes.

    https://doi.org/10.1136/jcp.52.10.766

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