A new fluorimetric method for the detection and quantification of siderophores using Calcein Blue, with potential as a bacterial detection tool

Verfasser / Beitragende:
[Ranjini Sankaranarayanan, Alagiachidambaram Alagumaruthanayagam, Krishnan Sankaran]
Ort, Verlag, Jahr:
2015
Enthalten in:
Applied Microbiology and Biotechnology, 99/5(2015-03-01), 2339-2349
Format:
Artikel (online)
ID: 60550430X
LEADER caa a22 4500
001 60550430X
003 CHVBK
005 20210128100615.0
007 cr unu---uuuuu
008 210128e20150301xx s 000 0 eng
024 7 0 |a 10.1007/s00253-015-6411-x  |2 doi 
035 |a (NATIONALLICENCE)springer-10.1007/s00253-015-6411-x 
245 0 2 |a A new fluorimetric method for the detection and quantification of siderophores using Calcein Blue, with potential as a bacterial detection tool  |h [Elektronische Daten]  |c [Ranjini Sankaranarayanan, Alagiachidambaram Alagumaruthanayagam, Krishnan Sankaran] 
520 3 |a 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. 
540 |a Springer-Verlag Berlin Heidelberg, 2015 
690 7 |a Bacteria  |2 nationallicence 
690 7 |a Calcein Blue  |2 nationallicence 
690 7 |a Fluorescence assay  |2 nationallicence 
690 7 |a Non-invasive  |2 nationallicence 
690 7 |a Siderophores  |2 nationallicence 
700 1 |a Sankaranarayanan  |D Ranjini  |u Centre for Biotechnology, Anna University, Sardar Patel road, Guindy, 600 025, Chennai, India  |4 aut 
700 1 |a Alagumaruthanayagam  |D Alagiachidambaram  |u Centre for Biotechnology, Anna University, Sardar Patel road, Guindy, 600 025, Chennai, India  |4 aut 
700 1 |a Sankaran  |D Krishnan  |u Centre for Biotechnology, Anna University, Sardar Patel road, Guindy, 600 025, Chennai, India  |4 aut 
773 0 |t Applied Microbiology and Biotechnology  |d Springer Berlin Heidelberg  |g 99/5(2015-03-01), 2339-2349  |x 0175-7598  |q 99:5<2339  |1 2015  |2 99  |o 253 
856 4 0 |u https://doi.org/10.1007/s00253-015-6411-x  |q text/html  |z Onlinezugriff via DOI 
898 |a BK010053  |b XK010053  |c XK010000 
900 7 |a Metadata rights reserved  |b Springer special CC-BY-NC licence  |2 nationallicence 
908 |D 1  |a research-article  |2 jats 
949 |B NATIONALLICENCE  |F NATIONALLICENCE  |b NL-springer 
950 |B NATIONALLICENCE  |P 856  |E 40  |u https://doi.org/10.1007/s00253-015-6411-x  |q text/html  |z Onlinezugriff via DOI 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Sankaranarayanan  |D Ranjini  |u Centre for Biotechnology, Anna University, Sardar Patel road, Guindy, 600 025, Chennai, India  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Alagumaruthanayagam  |D Alagiachidambaram  |u Centre for Biotechnology, Anna University, Sardar Patel road, Guindy, 600 025, Chennai, India  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Sankaran  |D Krishnan  |u Centre for Biotechnology, Anna University, Sardar Patel road, Guindy, 600 025, Chennai, India  |4 aut 
950 |B NATIONALLICENCE  |P 773  |E 0-  |t Applied Microbiology and Biotechnology  |d Springer Berlin Heidelberg  |g 99/5(2015-03-01), 2339-2349  |x 0175-7598  |q 99:5<2339  |1 2015  |2 99  |o 253