N. A. Os’kina, A. Avdalyan, D. Subbotin, A. Lazarev, A. Kel, N. Kushlinskii, M. Filipenko
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引用次数: 0
摘要
DNA结构改变的分子遗传学分析是当今临床研究的热点之一。由于DNA的交联和断裂,使用从福尔马林固定和石蜡包埋(FFPE)组织标本中获得的DNA尤其具有挑战性。我们使用两种内部方案和Qiagen (QIAamp DNA FFPE组织试剂盒)和Roche(高纯FFPET DNA分离试剂盒)在俄罗斯市场上推出并用于临床实践的商用试剂盒,对FFPE组织的DNA提取方法进行了比较分析。为了评估分离DNA的数量和质量,我们使用real-time PCR对DNA产率、抑制杂质含量和片段化程度进行评分。我们的发现可能对医学实验室进行体细胞突变测试以进行靶向治疗选择,以及专门从事肿瘤基因组结构研究的研究人员在患者预后和肿瘤细胞对治疗的敏感性或耐药性预测方面有用。
Comparison of Analytical Characteristics of Commercial and in-House Methods for DNA Isolation from Paraffin Histology Blocks
One of the hotspots in clinical research today is molecular genetic analysis of structural DNA alterations. Working with DNA obtained from formalin-fixed and paraffin-embedded (FFPE) tissue specimens is particularly challenging, due to cross-linking and fragmentation of DNA. We performed a comparative analysis of DNA extraction methods from FFPE tissue using two in-house protocols and Qiagen (QIAamp DNA FFPE Tissue Kit) and Roche (High Pure FFPET DNA Isolation Kit) commercial kits presented in the Russian market and used in clinical practice. To assess the quantity and quality of the isolated DNA, we used the real-time PCR to rate DNA yield, the inhibited impurity content and the degree of fragmentation. Our findings may be useful for a medical laboratory that performs testing of somatic mutations for the targeted therapy selection, and researchers who specialized in tumor genome structure studies in respect to patient prognosis and prediction of the sensitivity or resistance of tumor cells to therapy.