Ultra-rapid DNA analysis using HyBeacon™ probes and direct PCR amplification from saliva

被引：32

作者：

French, DJ ^{[1
]}

Archard, CL ^{[1
]}

Andersen, MT ^{[1
]}

McDowell, DG ^{[1
]}

机构：

[1] LGC Teddington Ltd, BioAnalyt Innovat, Teddington TW11 0LY, Middx, England

来源：

MOLECULAR AND CELLULAR PROBES | 2002年 / 16卷 / 05期

关键词：

fluorescent probes; LightCycler; single nucleoticle polymorphism; allele discrimination; pharmacogenetics; saliva;

D O I：

10.1006/mcpr.2002.0425

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

We describe a novel probe technology, termed HyBeacons(TM), which provides a new homogeneous method for fluorescence-based sequence detection and allele discrimination. Employing a single nucleotide polymorphism located in the N-acetyltransferase 2 gene as a model system, we demonstrate the utility of HyBeacon probes for rapid and reliable sequence analysis. We also demonstrate that homozygous and heterozygous samples may be accurately identified using a single HyBeacon oligonucleoticle. Polymorphic DNA sequences were detected and differentiated by real-time PCR and melt peak methodologies, without performing extraction of genomic DNA prior to target amplification. Employing a combination of homogeneous HyBeacon analysis, the rapid thermal cycling conditions of the LightCycler(TM) and direct amplification from saliva, allowed samples to be genotyped within 30 min. Such rapid non-invasive diagnostic technologies may permit 'point-of-care' genetic testing to be performed in hospitals and doctor's surgeries. (C) 2002 Elsevier Science Ltd. All rights reserved.

引用

页码：319 / 326

页数：8

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