Neoh S H, Brisco M J, Firgaira F A, Trainor K J, Turner D R, Morley A A
Department of Haematology and Genetic Pathology, Flinders University of South Australia, Flinders Medical Centre, Australia.
J Clin Pathol. 1999 Oct;52(10):766-9. doi: 10.1136/jcp.52.10.766.
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.
一种基于荧光共振能量转移（FRET）和实时聚合酶链反应（PCR）的快速方法被用于对112名个体进行血色素沉着症基因分型以及对134名个体进行凝血因子V基因分型。将结果与基于PCR产物限制性酶切的传统方法进行比较。在246名个体中，两种方法对244名个体的分型结果一致；对于另外两名个体，测序显示他们被标准方法错误地进行了基因分型，但FRET方法对其进行了正确的基因分型。使用FRET探针进行实时PCR分析的简便性、速度和准确性使其成为临床实验室对血色素沉着症和凝血因子V基因进行基因分型的首选方法。
