Development of an electrochemical Limulus amebocyte lysate assay technique for portable and highly sensitive endotoxin sensor |
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Authors: | Inoue Kumi Y Takahashi Satoko Ino Kosuke Shiku Hitoshi Matsue Tomokazu |
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Institution: | R&D Center of Excellence of Integrated Microsystems, Tohoku University, Aramaki, Aoba, Sendai, Japan. inoue@bioinfo.che.tohoku.ac.jp |
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Abstract: | Here, we report the development of an electrochemical detection method for endotoxin based on the Limulus amebocyte lysate (LAL) assay. A mixture of LAL reagent and endotoxin sample solution was incubated for 1 h. The endotoxin activated a cascade reaction of zymogens contained in the LAL to generate p-nitroaniline (pNA) which was then electrochemically detected by differential pulse voltammetry (DPV). The generated pNA gave a clear peak at -0.75 V vs. silver/silver chloride (Ag/AgCl), which increased with the concentration of endotoxin in the LAL assay solution. This DPV detection was performed using an electrode chip device fabricated from a diamond-like carbon-coated glass substrate. This chip device could detect as low as 10 endotoxin units l(-1) at room temperature within 1 h. This novel electrochemical method for the detection of endotoxin appears promising for the development of compact, low-cost and easy-to-use sensors for on-site monitoring of potentially contaminated medical supplies, including dialysis fluid, transplanted tissue and culture medium for assisted reproduction. |
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