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Analysis of pathogenic bacteria using exogenous volatile organic compound metabolites and optical sensor detection

机译：外源性挥发性有机化合物代谢物和光学传感器检测分析病原体细菌

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摘要

A novel, low-cost and simple method for the detection of pathogenic bacteria is proposed. The approach is based on the generation of an exogenous volatile organic compound (VOC) produced by the addition of an enzyme substrate to the bacterial sample. The generated VOC is then trapped in agarose gel allowing colour development to take place; visual detection is then possible by both the naked eye and by colorimetric analysis. Agarose gel has been evaluated as both a suitable VOC trapping matrix and host for the colour-generating reagents. This proof of concept method allowed for the discrimination between beta-glucosidase and beta-alanyl aminopeptidase producing bacteria. Enterococcus faecium and Klebsiella pneumoniae are both beta-glucosidase producers and generated a yellow colour within agarose gels upon enzymatic hydrolysis of 2-nitrophenyl-beta-D-glucoside. Pseudomonas aeruginosa is a known beta-alanyl aminopeptidase producer and was shown to hydrolyse the trifluoroacetic acid (TFA) salt of 3-amino-N-phenylpropanamide resulting in the development of an orange colour within agarose gels spiked with the sodium salt of 1,2-naphthoquinone-4-sulfonic acid. 3-Amino-N-phenylpropanamide (as its TFA salt) and 2-nitrophenyl-beta-D-glucoside concentrations of 20 mu g mL(-1) (or 72 mu mol L-1) and 100 mu g mL(-1) (or 332 mu mol L-1), respectively were the minimum quantities required for colour production following 18 h of incubation. The use of 3-amino-N-phenylpropanamide, TFA salt indicated that synthesised enzyme substrates can be tailor-made to liberate exogenous VOCs for colour generation.
机译：提出了一种新的，低成本和检测致病细菌的简单方法。该方法基于通过向细菌样品中加入酶底物而产生的外源性挥发性有机化合物（VOC）。然后将产生的VOC捕获在琼脂糖凝胶中，允许颜色开发发生;然后通过肉眼和比色分析可以通过视觉检测。已评估琼脂糖凝胶作为适当的VOC捕获基质和用于显色试剂的宿主。这种概念方法证明允许β-葡糖苷酶和β-丙氨酸氨基肽酶产生细菌之间的辨别。肠球菌粪便和Klebsiella Pneumoniae都是β-葡糖苷酶生产商，并在酶水解的2-硝基苯基-D-葡糖苷的酶水解后产生黄色的颜色。铜绿假单胞菌是一种已知的β-丙氨酸氨基肽酶生产商，并显示用于水解3-氨基-N-苯基丙酰胺的三氟乙酸（TFA）盐，导致琼脂糖凝胶内的橙色颜色掺入1,2的钠盐内的橙色。 - 萘醌-4-磺酸。 3-氨基-N-苯基丙烷（作为其TFA盐）和2-硝基苯基-D-葡糖苷浓度为20μgml（-1）（或72μmoll-1）和100μgml（-1 ）（或332μmolL-1）分别是孵育后18小时后产生颜色所需的最小量。使用3-氨基-N-苯基丙酰胺，TFA盐表明，合成的酶衬底可以量身定制，以释放出色的外源VOC。

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  • 来源
    《RSC Advances》 |2015年第20期|共6页
  • 作者

    Tait Emma; Stanforth Stephen P.; Reed Stephen; Perry John D.; Dean John R.;

  • 作者单位

    Northumbria Univ Dept Appl Sci Newcastle Upon Tyne NE1 8ST Tyne &

    Wear England;

    Northumbria Univ Dept Appl Sci Newcastle Upon Tyne NE1 8ST Tyne &

    Wear England;

    Northumbria Univ Dept Appl Sci Newcastle Upon Tyne NE1 8ST Tyne &

    Wear England;

    Freeman Rd Hosp Dept Microbiol Newcastle Upon Tyne NE7 7DN Tyne &

    Wear England;

    Northumbria Univ Dept Appl Sci Newcastle Upon Tyne NE1 8ST Tyne &

    Wear England;

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  • 正文语种 eng
  • 中图分类 化学;
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