PCR AMPLIFICATION FROM PARAFFIN-EMBEDDED TISSUES - EFFECTS OF FIXATIVE AND FIXATION TIME
PCR AMPLIFICATION FROM PARAFFIN-EMBEDDED TISSUES - EFFECTS OF FIXATIVE AND FIXATION TIME
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DOI：
10.1093/ajcp/95.2.117
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发表时间：
1991-02-01
影响因子：
3.5
通讯作者：
MANOS, MM
中科院分区：
文献类型：
--
作者：
GREER, CE;PETERSON, SL;MANOS, MM
聚合酶链式反应 (PCR) DNA 扩增方法是石蜡包埋组织 (PET) 回顾性分析的强大新工具。该技术提供了与遗传性疾病和传染病相关的核酸序列的灵敏且特异的检测。然而，PET 加工条件对于扩增的适用性各不相同。作者检查了 11 种固定剂在 3 个固定时间的效果。固定效果通过处理组织中的 DNA 作为模板扩增长度为 110 至 1,327 个碱基对的 DNA 片段的能力来测量。研究发现，用丙酮或 10% 缓冲中性福尔马林固定的样本最适合随后的 PCR 分析。第二组固定剂，包括 Zamboni 固定剂、Clarke 固定剂、多聚甲醛、福尔马林-乙醇-乙酸和 Methacarn，都会降低扩增效率。分别用 Carnoy's、Zenker's 或 Bouin's 处理的组织更不适合进行扩增分析。
The polymerase chain reaction (PCR) DNA amplification method is a powerful new tool for the retrospective analysis of paraffin-embedded tissue (PET). The technique has afforded the sensitive and specific detection of nucleic acid sequences associated with genetic and infectious diseases. However, PET processing conditions vary in their suitability for amplification. The authors have examined the effects of 11 fixatives at three fixation times. The effect of fixation was measured by the ability of the DNA in a treated tissue to serve as a template for the amplification of DNA fragments that ranged from 110 to 1,327 base pairs in length. Specimens fixed in acetone or 10% buffered neutral formalin were found to be best suited for subsequent analysis by PCR. A second group of fixatives, including Zamboni's, Clarke's, paraformaldehyde, formalin-alcohol-acetic acid, and methacarn, compromised amplification efficiency. Tissues treated with Carnoy's, Zenker's, or Bouin's, respectively, were even less desirable for amplification analysis.