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首页> 外文期刊>Applied Microbiology and Biotechnology >A new fluorimetric method for the detection and quantification of siderophores using Calcein Blue, with potential as a bacterial detection tool
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A new fluorimetric method for the detection and quantification of siderophores using Calcein Blue, with potential as a bacterial detection tool

机译：一种新的荧光测定法，使用钙黄绿素蓝检测和定量铁载体，具有潜在的细菌检测工具

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摘要

The presence of microorganisms in biological fluids like urine and blood is an indication of vulnerability to infections. Iron is one of the important micronutrients required for bacterial growth. In an iron-deficit environment, bacteria release high-affinity iron-chelating compounds called siderophores which can be used as non-invasive target molecules for the detection of such pathogens. However, only limited reagents and procedures are available to detect the presence of these organic molecules. The present study aims at detecting the presence of siderophores in the iron-depleted media, exploiting the reversible quenching of Calcein Blue and iron(III) complex. The fluorescence of Calcein Blue is known to be quenched in the presence of iron(III); if a stronger chelator removes this ion from the fluorophore, the fluorescence of the fluorophore is regained. This behaviour of the fluorophore was exploited to detect and quantify siderophores down to 50 and 800 nM equivalent of standard siderophore, deferroxamine mesylate (desferal) in Dulbecco's PBS and siderophore quantification (SPQ) medium, respectively. The siderophores released by pathogens, equivalent to standard desferal, were in the range of 1.29 to 5.00 mu M and those for non-pathogens were below 1.19 mu M. The simple, sensitive and cost-effective method performed in a 96-well plate was able to detect and quantify iron chelators within 7-8 h of incubation.
机译：尿液和血液等生物体液中微生物的存在表明容易感染。铁是细菌生长所需的重要微量营养素之一。在铁缺乏的环境中，细菌会释放出高亲和力的铁螯合化合物，称为铁载体，可以用作检测此类病原体的非侵入性目标分子。但是，只有有限的试剂和程序可用于检测这些有机分子的存在。本研究旨在通过利用钙黄绿素蓝和铁（III）络合物的可逆淬灭来检测铁耗尽介质中铁载体的存在。已知钙黄绿素蓝的荧光在铁（III）的存在下被猝灭。如果较强的螯合剂从荧光团上除去了该离子，则荧光团的荧光会重新获得。荧光团的这种行为被用来分别检测和定量低至50和800 nM当量的标准铁载体，Dulbecco PBS中的甲磺酸去铁胺（Serferal）和铁载体定量（SPQ）培养基。由病原体释放的铁载体相当于标准延迟，范围在1.29至5.00μM之间，非病原体的铁载体低于1.19μM。在96孔板上进行的简单，灵敏且经济高效的方法是能够在孵育的7-8小时内检测和定量铁螯合剂。

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