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Related Experiment Videos

Amphotericin B enzyme-linked immunosorbent assay

J D Cleary1, S W Chapman, J Deng

  • 1Department of Clinical Pharmacy, University of Mississippi Medical Center, Jackson, 39216-4505, USA.

Antimicrobial Agents and Chemotherapy
|March 1, 1996
PubMed
Summary
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A new enzyme-linked immunosorbent assay (ELISA) accurately measures amphotericin B in serum. This rapid assay aids research into antifungal drug efficacy and toxicity.

Area of Science:

  • * Clinical Chemistry
  • * Immunology
  • * Pharmacology

Background:

  • * Accurate measurement of antifungal drug serum concentrations is crucial for clinical research.
  • * Existing assays for amphotericin B are labor-intensive, limiting their use.

Purpose of the Study:

  • * To develop and characterize an enzyme-linked immunosorbent assay (ELISA) for quantifying amphotericin B in serum.
  • * To evaluate the assay's sensitivity, specificity, precision, and accuracy.

Main Methods:

  • * Competition ELISA using multiwell plates coated with amphotericin B-conjugated bovine serum albumin.
  • * Assay validation included testing sensitivity, specificity, precision, accuracy, and lipid-associated drug measurement.
  • * Analysis of reference samples and calculation of coefficients of variation and correlation coefficient.

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Main Results:

  • * The assay demonstrated a detection range of 0.15 to 156 micrograms/ml.
  • * Sensitivity was affected by light and temperature; specificity was only impacted by nystatin.
  • * Intrarun and interrun coefficients of variation were 3.0% and 12.8%, respectively, with a correlation coefficient of 0.907.

Conclusions:

  • * The developed ELISA is a precise, accurate, and easy-to-use method for measuring serum amphotericin B concentrations.
  • * This assay can facilitate clinical research by enabling rapid measurement of multiple samples.
  • * It addresses previous limitations posed by labor-intensive assays, aiding research into drug toxicity and efficacy.