Canzian F, Salovaara R, Hemminki A, Kristo P, Chadwick R B, Aaltonen L A, de la Chapelle A
Department of Medical Genetics, Haartman Institute, University of Helsinki, Finland.
Cancer Res. 1996 Jul 15;56(14):3331-7.
Loss of heterozygosity (LOH) and replication error (RER) are important phenomena in tumor development, with diagnostic and prognostic relevance. Therefore, screening for LOH and RER is a desirable first step in the molecular analysis of tumors. We used semiautomated procedures based on multicolor fluorescently labeled microsatellite markers and an automated sequencer for PCR amplification, electrophoresis of PCR products, and allele detection with a set of 16 microsatellites in 56 colorectal tumors. We improved existing software for computer-assisted assessment of LOH and RER. A comparison of these results with those of a conventional, radioactive technique and visual interpretation shows a high degree of correlation between the two methods. The detection rates of LOH and RER are similar to those reported previously. The main advantages of the semiautomated fluorescence-based typing are in the objective, observer-unrelated, easy, and rapid computer-based scoring, and the resulting quantitative assessment of RER.
杂合性缺失（LOH）和复制错误（RER）是肿瘤发生发展中的重要现象，具有诊断和预后相关性。因此，筛查LOH和RER是肿瘤分子分析中理想的第一步。我们使用基于多色荧光标记微卫星标记的半自动程序以及自动测序仪进行PCR扩增、PCR产物电泳，并使用一组16个微卫星对56例结直肠癌肿瘤进行等位基因检测。我们改进了现有的用于计算机辅助评估LOH和RER的软件。将这些结果与传统放射性技术及视觉判读的结果进行比较，显示两种方法之间具有高度相关性。LOH和RER的检测率与先前报道的相似。基于荧光的半自动分型的主要优点在于客观、与观察者无关、基于计算机的评分简便快速，以及由此对RER进行的定量评估。
