Rapid detection of the factor V Leiden (1691G>A) and haemochromatosis (845G>A) mutation by fluorescence resonance energy transfer (FRET) and real time PCR

S. H. Neoh; M. J. Brisco; F. A. Firgaira; K. J. Trainor; D. R. Turner; A. A. Morley

doi:10.1136/jcp.52.10.766

Rapid detection of the factor V Leiden (1691G>A) and haemochromatosis (845G>A) mutation by fluorescence resonance energy transfer (FRET) and real time PCR

S. H. Neoh, M. J. Brisco, F. A. Firgaira, K. J. Trainor, D. R. Turner, A. A. Morley

Research output: Contribution to journal › Article › peer-review

25 Citations (Scopus)

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.

Original language	English
Pages (from-to)	766-769
Number of pages	4
Journal	Journal of Clinical Pathology
Volume	52
Issue number	10
DOIs	https://doi.org/10.1136/jcp.52.10.766
Publication status	Published - Oct 1999

Keywords

Factor V Leiden
Fluorescence resonance energy transfer (FRET)
Haemochromatosis
LightCycler
Real time PCR

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title = "Rapid detection of the factor V Leiden (1691G>A) and haemochromatosis (845G>A) mutation by fluorescence resonance energy transfer (FRET) and real time PCR",

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.",

keywords = "Factor V Leiden, Fluorescence resonance energy transfer (FRET), Haemochromatosis, LightCycler, Real time PCR",

author = "Neoh, {S. H.} and Brisco, {M. J.} and Firgaira, {F. A.} and Trainor, {K. J.} and Turner, {D. R.} and Morley, {A. A.}",

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AU - Neoh, S. H.

AU - Brisco, M. J.

AU - Firgaira, F. A.

AU - Trainor, K. J.

AU - Turner, D. R.

AU - Morley, A. A.

PY - 1999/10

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N2 - 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.

AB - 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.

KW - Factor V Leiden

KW - Fluorescence resonance energy transfer (FRET)

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Rapid detection of the factor V Leiden (1691G>A) and haemochromatosis (845G>A) mutation by fluorescence resonance energy transfer (FRET) and real time PCR

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Keywords

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