Hukari Kyle W, Shultz Michael, Isely Nikolas, Milson Richard, West Jay A A
Arcxis Biotechnologies, Pleasanton, CA 94566, USA.
J Lab Autom. 2011 Oct;16(5):355-65. doi: 10.1016/j.jala.2010.08.004. Epub 2011 Mar 23.
Molecular diagnostic analysis and life science studies are dependent on the ability to effectively prepare samples for analysis. We report the development of a system that enables robust sample preparation of nucleic acids. To enable completely automated sample preparation, a consumable cartridge and consumable module system were developed to emulate every step of the sample preparation process. This included enzyme and reagent addition, temperature-controlled incubations, noncontact mixing of enzymes and reagents, buffer exchanges, and sample elution. Using this system, completely automated methods were developed for the purification of viral RNA and DNA from plasma and whole blood and of bacterial genomic DNA from water and whole blood. Extracted nucleic acids were detected and quantified using real-time PCR. The data indicate that automated viral DNA extraction was more efficient than sample extractions performed using a manual process, whereas automated total RNA extraction from the same samples was equivalent to controls. Additionally, we found that the process for bacterial genomic DNA extraction from either water or whole blood was equivalent to the manual extraction processes. We conclude the instrument, consumable cartridge, and reagent system enables easy, cost-effective, and robust sample preparation regardless of the experience of the operator.
分子诊断分析和生命科学研究依赖于有效制备分析样本的能力。我们报告了一种能够对核酸进行可靠样本制备的系统的开发情况。为实现完全自动化的样本制备，开发了一种消耗品盒和消耗品模块系统，以模拟样本制备过程的每一步。这包括添加酶和试剂、控温孵育、酶和试剂的非接触混合、缓冲液交换以及样本洗脱。利用该系统，开发了从血浆和全血中纯化病毒RNA和DNA以及从水和全血中纯化细菌基因组DNA的完全自动化方法。使用实时PCR对提取的核酸进行检测和定量。数据表明，自动化病毒DNA提取比使用手动方法进行的样本提取更有效，而从相同样本中进行的自动化总RNA提取与对照相当。此外，我们发现从水或全血中提取细菌基因组DNA的过程与手动提取过程相当。我们得出结论，该仪器、消耗品盒和试剂系统能够实现简便、经济高效且可靠的样本制备，而无需考虑操作人员的经验。
