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Rapid Universal Solublization and Analysis of bacterial spores using a Simple Flow-Through Ultra-High Temperature Capillary Device

机译：使用简单的流通式超高温毛细管装置快速通用溶解和分析细菌孢子

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摘要

Rapid identification of viral and bacterial species is dependent of the ability to manipulate biological agents into a form where they are directly analyzed. Many of these species of interest, such as bacterial spores, are inherently hearty and very difficult to lyse or solubilize. Standard protocols for spore inactivation include, chemical treatment, sonication, pressure and thermal lysis. While these protocols are effective for the inactivation of these agents they are less well suited for sample preparation for analysis using capillary electrophoresis techniques. In order to overcome this difficulty we fabricated a simple capillary device to perform thermal lysis of vegatative bacterial cells and bacterial spores. Using an ethylene glycol buffer to super heat bacterial spores we were able to perform rapid flow through lysis and solubilzation of these agents. This device was then coupled to a sample preparation station for on-line fluorescamine dye lableling and buffer exchange for direct analysis using a miniaturized capillary electrophoresis instrument. Using this integrated device were we enabled to perform sample lysis, labeling and protein fingerprint analysis of vegatative bacterial cells, bacterial spores and viruses in less than 10 minutes. The described device is simple, inexpensive and easily integratable with various microfluidic devices.
机译：快速鉴定病毒和细菌种类取决于将生物制剂操纵为直接对其进行分析的形式的能力。这些感兴趣的物种中的许多物种（例如细菌孢子）天生就具有丰盛性，很难溶解或溶解。孢子灭活的标准方案包括化学处理，超声处理，压力和热裂解。尽管这些方案对于这些试剂的灭活是有效的，但它们不太适合使用毛细管电泳技术进行分析的样品制备。为了克服这一困难，我们制造了一种简单的毛细管装置，用于对溶菌性细菌细胞和细菌孢子进行热裂解。使用乙二醇缓冲液使细菌孢子过热，我们能够通过裂解和溶解这些试剂来实现快速流动。然后将此设备连接到样品制备站，以使用微型毛细管电泳仪在线进行荧光胺染料标记和缓冲液交换，以进行直接分析。使用该集成设备，我们能够在不到10分钟的时间内对变性细菌细胞，细菌孢子和病毒进行样品裂解，标记和蛋白质指纹分析。所描述的设备简单，便宜并且容易与各种微流体设备集成。

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