caa a22 4500 60550430X CHVBK 20210128100615.0 cr unu---uuuuu 210128e20150301xx s 000 0 eng 10.1007/s00253-015-6411-x doi (NATIONALLICENCE)springer-10.1007/s00253-015-6411-x A new fluorimetric method for the detection and quantification of siderophores using Calcein Blue, with potential as a bacterial detection tool [Elektronische Daten] [Ranjini Sankaranarayanan, Alagiachidambaram Alagumaruthanayagam, Krishnan Sankaran] 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 800nM 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 5.00μM and those for non-pathogens were below 1.19μM. The simple, sensitive and cost-effective method performed in a 96-well plate was able to detect and quantify iron chelators within 7-8h of incubation. Springer-Verlag Berlin Heidelberg, 2015 Bacteria nationallicence Calcein Blue nationallicence Fluorescence assay nationallicence Non-invasive nationallicence Siderophores nationallicence Sankaranarayanan Ranjini Centre for Biotechnology, Anna University, Sardar Patel road, Guindy, 600 025, Chennai, India aut Alagumaruthanayagam Alagiachidambaram Centre for Biotechnology, Anna University, Sardar Patel road, Guindy, 600 025, Chennai, India aut Sankaran Krishnan Centre for Biotechnology, Anna University, Sardar Patel road, Guindy, 600 025, Chennai, India aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/5(2015-03-01), 2339-2349 0175-7598 99:5<2339 2015 99 253 https://doi.org/10.1007/s00253-015-6411-x text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-015-6411-x text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Sankaranarayanan Ranjini Centre for Biotechnology, Anna University, Sardar Patel road, Guindy, 600 025, Chennai, India aut NATIONALLICENCE 700 1- Alagumaruthanayagam Alagiachidambaram Centre for Biotechnology, Anna University, Sardar Patel road, Guindy, 600 025, Chennai, India aut NATIONALLICENCE 700 1- Sankaran Krishnan Centre for Biotechnology, Anna University, Sardar Patel road, Guindy, 600 025, Chennai, India aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/5(2015-03-01), 2339-2349 0175-7598 99:5<2339 2015 99 253