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Evaluation of ligand modified poly (N-Isopropyl acrylamide) hydrogel for etiological diagnosis of corneal infectionShivshetty, N., Swift, Thomas, Pinnock, A., Pownall, D., MacNeil, S., Douglas, I., Garg, P., Rimmer, Stephen 24 March 2022 (has links)
Yes / Corneal ulcers, a leading cause of blindness in the developing world are treated inappropriately without prior
microbiology assessment because of issues related to availability or cost of accessing these services.
In this work we aimed to develop a device for identifying the presence of Gram-positive or Gram-negative
bacteria or fungi that can be used by someone without the need for a microbiology laboratory. Working with
branched poly (N-isopropyl acrylamide) (PNIPAM) tagged with Vancomycin, Polymyxin B, or Amphotericin B to
bind Gram-positive bacteria, Gram-negative bacteria and fungi respectively, grafted onto a single hydrogel we
demonstrated specific binding of the organisms. The limit of detection of the microbes by these polymers was
between 10 and 4 organisms per high power field (100X) for bacteria and fungi binding polymers respectively.
Using ex vivo and animal cornea infection models infected with bacteria, fungi or both we than demonstrated
that the triple functionalised hydrogel could pick up all 3 organisms after being in place for 30 min. To confirm
the presence of bacteria and fungi we used conventional microbiology techniques and fluorescently labelled
ligands or dyes.
While we need to develop an easy-to-use either a colorimetric or an imaging system to detect the fluorescent
signals, this study presents for the first time a simple to use hydrogel system, which can be applied to infected
eyes and specifically binds different classes of infecting agents within a short space of time. Ultimately this
diagnostic system will not require trained microbiologists for its use and will be used at the point-of-care. / We gratefully acknowledge support for this research by the Well- come Trust which provided funding for Shivshetty, Swift and Pinnock (Grant 0998800/B/12/Z).
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