快速自动DNA和RNA提取技术在癌症诊断中检测体细胞或体质基因异常的比较

F. Lassailly, M. Mozziconacci, T. Noguchi, Didier Bechlian, J. Adélaı̈de, F. Fina, P. Martin, H. Sobol, L. Xerri, D. Birnbaum, C. Chabannon
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引用次数: 2

摘要

大量患者样本的分子分析越来越多地用于诊断应用以及了解癌症生物学。它们的准确性取决于从人体细胞或组织中提取的核酸的质量和数量。为了优化这些分析前步骤,我们评估了几种用于临床样品核酸纯化的自动化技术。对三种自动化平台进行了比较。从5名正常人的外周血白细胞中提取DNA,采用D-HPLC法和PCR测序法评价其质量。采用急性白血病患者临床标本进行RNA自动提取;结果与我们的标准手工技术进行了比较。用毛细管凝胶电泳鉴定RNA,用实时荧光定量PCR分析Abelson基因转录物。一个机器人对DNA和RNA的总产出更高。虽然从三个工作站获得的dna质量允许实施他们的分析…
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Comparison of a Selection of Rapid Automated DNA and RNA Extraction Technologies for Detection of Somatic or Constitutional Gene Abnormalities in Cancer Diagnosis
Molecular analyses of large numbers of patient samples are increasingly used for diagnostic applications as well as for understanding cancer biology. Their accuracy depends on the quality and quantity of nucleic acids extracted from human cells or tissues. To optimize these preanalytical steps, we evaluated several automated technologies for nucleic acid purification from clinical samples. Three automated platforms were compared. DNA was extracted from peripheral blood leukocytes from five normal individuals, and its quality was assessed by D-HPLC and sequencing after PCR. Clinical samples from acute leukemia patients were used for automated RNA extractions; results were compared to our standard manual technique. RNA qualification was done using capillary gel electrophoresis and analysis of the Abelson gene transcript by real-time PCR. One robot produced higher total output for both DNA and RNA. While the quality of DNAs obtained from the three workstations allowed implementation of their analysis for det...
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