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

被引:22
作者
Neoh, SH [1 ]
Brisco, MJ [1 ]
Firgaira, FA [1 ]
Trainor, KJ [1 ]
Turner, DR [1 ]
Morley, AA [1 ]
机构
[1] Flinders Univ S Australia, Flinders Med Ctr, Dept Haematol & Gen Pathol, Bedford Pk, SA 5042, Australia
关键词
real rime PCR; LightCycler; fluorescence resonance energy transfer (FRET); factor V Leiden; haemochromatosis;
D O I
10.1136/jcp.52.10.766
中图分类号
R36 [病理学];
学科分类号
100104 ;
摘要
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.
引用
收藏
页码:766 / 769
页数:4
相关论文
共 10 条
[1]  
BALNAVES ME, 1991, NUCLEIC ACIDS RES, V19, P2537
[2]   RAPID 2-STAGE PCR FOR DETECTING FACTOR-V G1691A-MUTATION [J].
BEAUCHAMP, NJ ;
DALY, ME ;
COOPER, PC ;
PRESTON, FE ;
PEAKE, IR .
LANCET, 1994, 344 (8923) :694-695
[3]   Homogeneous multiplex genotyping of hemochromatosis mutations with fluorescent hybridization probes [J].
Bernard, PS ;
Ajioka, RS ;
Kushner, JP ;
Wittwer, CT .
AMERICAN JOURNAL OF PATHOLOGY, 1998, 153 (04) :1055-1061
[4]   MUTATION IN BLOOD-COAGULATION FACTOR-V ASSOCIATED WITH RESISTANCE TO ACTIVATED PROTEIN-C [J].
BERTINA, RM ;
KOELEMAN, BPC ;
KOSTER, T ;
ROSENDAAL, FR ;
DIRVEN, RJ ;
DERONDE, H ;
VANDERVELDEN, PA ;
REITSMA, PH .
NATURE, 1994, 369 (6475) :64-67
[5]   A novel MHC class I-like gene is mutated in patients with hereditary haemochromatosis [J].
Feder, JN ;
Gnirke, A ;
Thomas, W ;
Tsuchihashi, Z ;
Ruddy, DA ;
Basava, A ;
Dormishian, F ;
Domingo, R ;
Ellis, MC ;
Fullan, A ;
Hinton, LM ;
Jones, NL ;
Kimmel, BE ;
Kronmal, GS ;
Lauer, P ;
Lee, VK ;
Loeb, DB ;
Mapa, FA ;
McClelland, E ;
Meyer, NC ;
Mintier, GA ;
Moeller, N ;
Moore, T ;
Morikang, E ;
Prass, CE ;
Quintana, L ;
Starnes, SM ;
Schatzman, RC ;
Brunke, KJ ;
Drayna, DT ;
Risch, NJ ;
Bacon, BR ;
Wolff, RK .
NATURE GENETICS, 1996, 13 (04) :399-408
[6]   Haemochromatosis and HLA-H [J].
Jouanolle, AM ;
Gandon, G ;
Jezequel, P ;
Blayau, M ;
Campion, ML ;
Yaouanq, J ;
Mosser, J ;
Fergelot, P ;
Chauvel, B ;
Bouric, P ;
Carn, G ;
Andrieux, N ;
Gicquel, I ;
LeGall, JY ;
David, V .
NATURE GENETICS, 1996, 14 (03) :251-252
[7]   Detection of the hereditary hemochromatosis gene mutation by real-time fluorescence polymerase chain reaction and peptide nucleic acid clamping [J].
Kyger, EM ;
Krevolin, MD ;
Powell, MJ .
ANALYTICAL BIOCHEMISTRY, 1998, 260 (02) :142-148
[8]  
Lay MJ, 1997, CLIN CHEM, V43, P2262
[9]   The LightCycler(TM) a microvolume multisample fluorimeter with rapid temperature control [J].
Wittwer, CT ;
Ririe, KM ;
Andrew, RV ;
David, DA ;
Gundry, RA ;
Balis, UJ .
BIOTECHNIQUES, 1997, 22 (01) :176-181
[10]   LINKAGE BETWEEN INHERITED RESISTANCE TO ACTIVATED PROTEIN-C AND FACTOR-V GENE MUTATION IN VENOUS THROMBOSIS [J].
ZOLLER, B ;
DAHLBACK, B .
LANCET, 1994, 343 (8912) :1536-1538