Weiss M M, Hermsen M A, Meijer G A, van Grieken N C, Baak J P, Kuipers E J, van Diest P J
Department of Gastroenterology, Free University Hospital, Amsterdam, The Netherlands.
Mol Pathol. 1999 Oct;52(5):243-51. doi: 10.1136/mp.52.5.243.
Comparative genomic hybridisation (CGH) is a technique that permits the detection of chromosomal copy number changes without the need for cell culturing. It provides a global overview of chromosomal gains and losses throughout the whole genome of a tumour. Tumour DNA is labelled with a green fluorochrome, which is subsequently mixed (1:1) with red labelled normal DNA and hybridised to normal human metaphase preparations. The green and red labelled DNA fragments compete for hybridisation to their locus of origin on the chromosomes. The green to red fluorescence ratio measured along the chromosomal axis represents loss or gain of genetic material in the tumour at that specific locus. In addition to a fluorescence microscope, the technique requires a computer with dedicated image analysis software to perform the analysis. This review aims to provide a detailed discussion of the CGH technique, and to provide a protocol with an emphasis on crucial steps.
比较基因组杂交(CGH)是一种无需细胞培养就能检测染色体拷贝数变化的技术。它能全面概述肿瘤整个基因组中的染色体增减情况。肿瘤DNA用绿色荧光染料标记,随后与红色标记的正常DNA按1:1混合,并与正常人中期染色体标本杂交。绿色和红色标记的DNA片段竞争与染色体上其起源位点的杂交。沿染色体轴测量的绿色与红色荧光比率代表肿瘤在该特定位点的遗传物质丢失或增加。除了荧光显微镜外,该技术还需要一台装有专用图像分析软件的计算机来进行分析。本综述旨在详细讨论CGH技术,并提供一个强调关键步骤的方案。