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Fluorescence polarization immunoassay for zidovudine
G G Granich1, M R Eveland, D J Krogstad
1Department of Medicine, Washington University School of Medicine, St. Louis, Missouri.
Antimicrobial Agents and Chemotherapy
|August 1, 1989
Summary
A new fluorescence polarization immunoassay (FPIA) accurately measures zidovudine (azidothymidine) in patient serum. This sensitive and specific assay is faster than HPLC and useful for HIV clinical trials.
Area of Science:
- Clinical Chemistry
- Immunochemistry
- Pharmacokinetics
Background:
- Zidovudine (azidothymidine) is a critical antiretroviral medication.
- Accurate therapeutic drug monitoring of zidovudine is essential for patient management.
- Existing analytical methods may have limitations in sensitivity, specificity, or throughput.
Purpose of the Study:
- To develop and validate a fluorescence polarization immunoassay (FPIA) for quantifying zidovudine in human serum.
- To assess the assay's accuracy, specificity, and performance compared to existing methods.
- To evaluate the assay's utility in clinical settings, particularly for large patient cohorts.
Main Methods:
- Development of a fluorescence polarization immunoassay (FPIA) for zidovudine.
- Validation of the assay across a clinically relevant concentration range (1-1,250 ng/ml).
- Comparison of FPIA with high-performance liquid chromatography (HPLC) and radioimmunoassay (RIA).
Main Results:
- The FPIA demonstrated high accuracy and specificity, unaffected by common interfering substances in patient serum.
- Cross-reactivity with zidovudine analogs was minimal (<0.05% for most).
- FPIA showed superior sensitivity, required less sample volume, and was significantly faster than HPLC (45 vs. 3 specimens/h).
- FPIA correlated highly with RIA (r=0.992).
- All tested assays were unaffected by heat treatment for HIV inactivation.
Conclusions:
- The developed FPIA is a sensitive, specific, and rapid method for zidovudine measurement.
- This assay offers significant advantages over HPLC for routine clinical laboratory use.
- The FPIA is well-suited for monitoring zidovudine therapy in large-scale clinical trials and patient populations.