Akonni Biosystems, Inc., Frederick, MD 21701, USA.
J Virol Methods. 2012 Jul;183(1):8-13. doi: 10.1016/j.jviromet.2012.03.002. Epub 2012 Mar 7.
This report describes the development and pre-clinical testing of a new, random-access RNA sample preparation system (TruTip) for nasopharyngeal samples. The system is based on a monolithic, porous nucleic acid binding matrix embedded within an aerosol-resistant pipette tip and can be operated with single or multi-channel pipettors. Equivalent extraction efficiencies were obtained between automated QIAcube and manual TruTip methods at 10(6) gene copies influenza A per mL nasopharyngeal aspirate. Influenza A and B amended into nasopharyngeal swabs (in viral transport medium) were detected by real-time RT-PCR at approximately 745 and 370 gene copies per extraction, respectively. RNA extraction efficiency in nasopharyngeal swabs was also comparable to that obtained on an automated QIAcube instrument over a range of input concentrations; the correlation between threshold cycles (or nucleic acid recovery) for TruTip and QIAcube-purified RNA was R(2)>0.99. Preclinical testing of TruTip on blinded nasopharyngeal swab samples resulted in 98% detection accuracy relative to a clinically validated easyMAG extraction method. The physical properties of the TruTip binding matrix and ability to customize its shape and dimensions likewise make it amenable to automation and/or fluidic integration.
本报告描述了一种新的、随机访问的 RNA 样品制备系统（TruTip）的开发和临床前测试，该系统基于嵌入在耐气溶胶移液器吸头内的整体式多孔核酸结合基质，可与单通道或多通道移液器一起使用。在 10（6）个基因拷贝/毫升鼻咽抽吸物中，自动 QIAcube 和手动 TruTip 方法之间获得了相当的提取效率。用实时 RT-PCR 检测到在病毒运输培养基中加入到鼻咽拭子中的甲型和乙型流感，分别提取约 745 和 370 个基因拷贝。在鼻咽拭子中，RNA 提取效率也与在自动化 QIAcube 仪器上获得的效率相当，在输入浓度范围内，TruTip 和 QIAcube 纯化 RNA 的阈值循环（或核酸回收率）之间的相关性 R（2）>0.99。在盲法鼻咽拭子样本上对 TruTip 的临床前测试，其检测准确率相对于经过临床验证的 easyMAG 提取方法为 98%。TruTip 结合基质的物理特性及其定制形状和尺寸的能力，使其同样适用于自动化和/或流体集成。
