LightCycler Technology in Molecular Diagnostics
LightCycler Technology in Molecular Diagnostics
复制标题
DOI：
10.2353/jmoldx.2009.080094
复制
发表时间：
2009-03-01
影响因子：
4.1
通讯作者：
Wittwer, Carl T.
中科院分区：
文献类型：
--
作者：
Lyon, Elaine;Wittwer, Carl T.
LightCycler 技术将快速循环聚合酶链反应与实时荧光监测和熔解曲线分析相结合。自1997年推出以来，现已应用于分子病理学的许多领域，包括肿瘤学（实体瘤和血液病理学）、遗传性疾病和传染病。通过监测快速扩增过程中产物的积累，可以在 15 至 30 分钟内实现闭管系统中的定量聚合酶链反应。此外，探针和/或扩增子的熔解曲线分析提供了基因分型甚至单倍型分析。新的突变是通过意想不到的解链温度或曲线形状变化来识别的。熔解探针设计包括相邻杂交探针、单标记探针、未标记探针和折回引物。高分辨率熔解允许通过检测所有杂合变化来进行突变扫描。本综述描述了过去 1.5 年有关 LightCycler 技术及其在临床实验室中的应用的主要进展。(j Moldx Diagn 2009, 11-9.3-101; DOI: 10.23531/jmoldx.2009.080094)
LightCycler technology combines rapid-cycle polymerase chain reaction with real-time fluorescent monitoring and melting curve analysis. Since its introduction in 1997, It is now used in many areas of molecular pathology, including oncology (solid tumors and hematopathology), inherited disease, and infectious disease. By monitoring product accumulation during rapid amplification, quantitative polymerase chain reaction in a closed-tube system is possible in 15 to 30 minutes. Furthermore, melting curve analysis of probes and/or amplicons provides genotyping and even haplotyping. Novel mutations are identified by unexpected melting temperature or curve shape changes. Melting probe designs include adjacent hybridization probes, single labeled probes, unlabeled probes, and snapback primers. High-resolution melting allows mutation scanning by detecting all heterozygous changes. This review describes the major advances throughout the last 1.5 years regarding LightCycler technology and its application in clinical laboratories.(j Moldx Diagn 2009, 11-9.3-101; DOI: 10.23531/jmoldx.2009.080094)