Antibiotic susceptibility testing at a screen-printed carbon electrode array

Antibiotic susceptibility testing at a screen-printed carbon electrode array
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DOI:
10.1021/ac701829c
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发表时间:
2008-02-01
影响因子:
7.4
通讯作者:
Mikkelsen, Susan R.
Mikkelsen, Susan R.
中科院分区:
化学1区
文献类型:
--
作者:
Mann, Thomas S.;Mikkelsen, Susan R.

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对丝网印刷碳电极阵列进行处理,以进行基于呼吸活动的测量。大肠埃希菌JM105对抗生素敏感性的研究研究了碳工作电极对抗生素的吸附,并用不同的电化学和化学方案对其进行了预处理,以最大限度地减少抗生素的吸附。在碱性溶液中用伏安法处理或用聚L赖氨酸或壳聚糖化学修饰是消除17种抗生素吸附的有效方法,这些抗生素包括几类和几种作用方式。测量包括在与抗生素孵育10分钟后用双电极安培法测定细菌的悬浮液,然后添加铁氰化物和二氯苯酚的氧化鸡尾酒再作用10分钟;在最初的10分钟孵育中,指示细胞呼吸活性将铁氰化物还原为亚铁氰化物的程度的响应电流随着抗菌剂浓度的增加而减小。用这些电极修饰方法获得的氯霉素的IC50值在2.0+/-0.2 mm处一致,与以前报道的基于呼吸的结果大致一致;对于这种微生物,IC50值明显高于基于生长的抗生素敏感性测试法报告的值。
Screen-printed carbon electrode arrays were treated to allow respiratory activity-based measurement. of antibiotic susceptibility with Escherichia coli JM105. Carbon working electrodes were examined for antibiotic adsorption and were pretreated with various electrochemical and chemical protocols to minimize 'antibiotic adsorption. Treatment by voltammetry in basic solution or by chemical modification with poly-L-lysine or chitosan were found to be effective methods for the elimination of adsorption of the, studied group of 17 antibiotics, which comprised several classes and modes of action. Measurements consisted of two-Electrode amperometry of the bacteria] suspension after 10 min of incubation with antibiotic followed by addition of an oxidative cocktail of ferricyanide and dichlorophenolindophenol for a further 10 min; response currents, which indicate the extent of reduction of ferricyanide to ferrocyanide by cellular respiratory activity, decrease with increasing concentration of anti-biotic present in the initial 10 min incubation. IC50 values obtained for chloramphenicol with these electrode modification methods are consistent at 2.0 +/- 0.2 mM, in approximate agreement with previously reported respiration-based results; for this organism but significantly higher than values reported for growth-based antibiotic susceptibility testing methods.