Dielectrophoretic monitoring and interstrain separation of intact Clostridium difficile based on their S(Surface)-layers.
Dielectrophoretic monitoring and interstrain separation of intact Clostridium difficile based on their S(Surface)-layers.
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DOI：
10.1021/ac5029837
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发表时间：
2014-11-04
影响因子：
7.4
通讯作者：
Swami, Nathan S.
中科院分区：
文献类型：
--
作者：
Su, Yi-Hsuan;Warren, Cirle A.;Guerrant, Richard L.;Swami, Nathan S.
艰难梭菌（C. difficile）感染（CDI）率在过去二十年中在世界范围内呈稳步上升趋势，由于抗生素耐药菌株的出现，感染构成全球威胁。跨宿主微生物组的菌株间拮抗相互作用形成了控制CDI出现的重要策略。目前CDI的诊断方法是基于对致病性艰难梭菌菌株产生的毒素进行免疫测定，由于毒素降解迅速，因此存在假阴性的限制。此外，由于缺乏这些毒素，同时监测非产毒艰难梭菌菌株是不可能的，从而限制了其通过优化拮抗菌间相互作用来控制CDI的应用。在此，我们证明了具有不同s层蛋白的特定艰难梭菌菌株细胞壁内的形态差异可以引起其电生理的系统性变化，这是由于细胞壁电容的改变。因此，介电泳频率分析可以实现艰难梭菌混合样品中各菌株类型完整微生物的独立指纹和无标记分离。这种无接触电生理方法的灵敏度与免疫测定和微生物生长速率方法进行基准测试，用于检测万古霉素治疗后产毒和非产毒艰难梭菌菌株的变化。这种微流控诊断平台可以通过分离单个菌株、优化抗生素治疗和监测微生物组来帮助开发阻止梭状芽胞杆菌感染的治疗方法。
Clostridium difficile (C. difficile) infection (CDI) rates have exhibited a steady rise worldwide over the last two decades and the infection poses a global threat due to the emergence of antibiotic resistant strains. Interstrain antagonistic interactions across the host microbiome form an important strategy for controlling the emergence of CDI. The current diagnosis method for CDI, based on immunoassays for toxins produced by pathogenic C. difficile strains, is limited by false negatives due to rapid toxin degradation. Furthermore, simultaneous monitoring of nontoxigenic C. difficile strains is not possible, due to absence of these toxins, thereby limiting its application toward the control of CDI through optimizing antagonistic interstrain interactions. Herein, we demonstrate that morphological differences within the cell wall of particular C. difficile strains with differing S-layer proteins can induce systematic variations in their electrophysiology, due alterations in cell wall capacitance. As a result, dielectrophoretic frequency analysis can enable the independent fingerprinting and label-free separation of intact microbials of each strain type from mixed C. difficile samples. The sensitivity of this contact-less electrophysiological method is benchmarked against the immunoassay and microbial growth rate methods for detecting alterations within both, toxigenic and nontoxigenic C. difficile strains after vancomycin treatment. This microfluidic diagnostic platform can assist in the development of therapies for arresting clostridial infections by enabling the isolation of individual strains, optimization of antibiotic treatments and the monitoring of microbiomes.
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