Longy M, Duboue B, Soubeyran P, Moynet D
Laboratory of Molecular Oncology, Institut Bergonie, Bordeaux, France.
Diagn Mol Pathol. 1997 Jun;6(3):167-73. doi: 10.1097/00019606-199706000-00007.
Paraffin-embedded tissues are often the only available material to perform polymerase chain reaction (PCR)-based analysis in various medical purposes. Unfortunately, the use in many countries of acid fixatives such as Bouin's fluid limits the use of such a material for molecular analysis. This article reports the methodological details of a DNA purification technique from Bouin-fixed and paraffin-embedded samples based on a double washing, in an alcohol then in an aqueous medium, of the DNA, which enables PCR reactions from this material. Comparison of the results with those obtained by organic solvent purification of DNA from frozen tissue fragments showed excellent reproducibility in terms of detection of an amplification product on agarose gel. However, differences between the methods were quite frequently seen in the allelic typing profile of microsatellite sequences (CA repeats), either as neo-alleles or by the loss of normal alleles in the fixed materials that constitute a limitation in using DNA from Bouin-fixed tissue as a substrate for fine allelotyping.
石蜡包埋组织往往是出于各种医学目的进行基于聚合酶链反应（PCR）分析时唯一可用的材料。不幸的是，在许多国家，诸如Bouin氏液等酸性固定剂的使用限制了这种材料在分子分析中的应用。本文报告了一种从Bouin固定和石蜡包埋样本中纯化DNA的技术方法细节，该方法基于对DNA进行酒精然后水性介质的双重洗涤，从而能够对这种材料进行PCR反应。将结果与通过有机溶剂从冷冻组织片段中纯化DNA所获得的结果进行比较，发现在琼脂糖凝胶上检测扩增产物方面具有出色的重现性。然而，在微卫星序列（CA重复）的等位基因分型谱中，这两种方法之间经常出现差异，表现为新等位基因的出现，或者在固定材料中正常等位基因的丢失，这构成了将Bouin固定组织的DNA用作精细等位基因分型底物的一个限制因素。
