Soukup J, Krskova L, Hilska I, Kodet R
Laboratory of Molecular Pathology, Department of Pathology and Molecular Medicine, Charles University, 2nd Medical School, Prague 5 - Motol, 150 06 Czech Republic.
Neoplasma. 2003;50(4):300-4.
Molecular methods play an important role in diagnostic pathology of lymphomas. PCR based demonstration of clonality or detection of a specific chromosomal translocation may determine the exact classification of the lymphoma. Hence the final diagnosis may depend on the quality of preserved nucleic acids in the bioptic specimen. The integrity of DNA and RNA may be damaged by formalin fixation, which destroys the nucleic acids by fragmentation. Therefore, a portion of each lymphoma sample should be frozen. To substitute freezing techniques we utilized ethanol as a fixative, which preserves nucleic acids. We compared PCR and RT-PCR products from lymphoma samples, which were differently pre-treated by ethanol fixation, formalin fixation and freezing. The ethanol fixed samples retained a high quality of both DNA and RNA and provided reproducible PCR products similar to frozen samples and significantly better then those extracted from formalin fixed samples. We may recommend ethanol as a complementary fixative for all pathology laboratories where deep freezing in not routinely available.
分子方法在淋巴瘤的诊断病理学中发挥着重要作用。基于聚合酶链反应（PCR）的克隆性证明或特定染色体易位的检测可确定淋巴瘤的确切分类。因此，最终诊断可能取决于活检标本中保存的核酸质量。福尔马林固定会破坏核酸，导致DNA和RNA的完整性受损。因此，每个淋巴瘤样本都应冷冻一部分。为了替代冷冻技术，我们使用乙醇作为固定剂，它可以保存核酸。我们比较了淋巴瘤样本经乙醇固定、福尔马林固定和冷冻等不同预处理后的PCR和逆转录PCR（RT-PCR）产物。乙醇固定的样本保留了高质量的DNA和RNA，并提供了与冷冻样本相似的可重复PCR产物，且明显优于从福尔马林固定样本中提取的产物。对于所有无法常规进行深度冷冻的病理实验室，我们推荐使用乙醇作为补充固定剂。
