Lorenz Michael G, Disqué Claudia, Mühl Helge
Molzym GmbH & Co. KG, Mary-Astell-Straße 10, 28359, Bremen, Germany,
Methods Mol Biol. 2015;1237:121-8. doi: 10.1007/978-1-4939-1776-1_12.
Automation in DNA isolation is a necessity for routine practice employing molecular diagnosis of infectious agents. To this end, the development of automated systems for the molecular diagnosis of microorganisms directly in blood samples is at its beginning. Important characteristics of systems demanded for routine use include high recovery of microbial DNA, DNA-free containment for the reduction of DNA contamination from exogenous sources, DNA-free reagents and consumables, ideally a walkaway system, and economical pricing of the equipment and consumables. Such full automation of DNA extraction evaluated and in use for sepsis diagnostics is yet not available. Here, we present protocols for the semiautomated isolation of microbial DNA from blood culture and low- and high-volume blood samples. The protocols include a manual pretreatment step followed by automated extraction and purification of microbial DNA.
在采用传染病原体分子诊断的常规实践中,DNA提取自动化是必要的。为此,直接对血液样本中的微生物进行分子诊断的自动化系统尚处于起步阶段。常规使用所需系统的重要特性包括微生物DNA的高回收率、用于减少外源DNA污染的无DNA密闭环境、无DNA试剂和耗材、理想情况下的自动运行系统,以及设备和耗材的经济定价。用于脓毒症诊断的DNA提取全自动化评估及应用目前尚不可用。在此,我们展示了从血培养以及少量和大量血液样本中半自动化分离微生物DNA的方案。这些方案包括一个手动预处理步骤,随后是微生物DNA的自动提取和纯化。