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Multiplexed Fluorometric ImmunoAssay Testing Methodology and Troubleshooting
Published on: December 12, 2011
Assay of teicoplanin in serum: comparison of high-performance liquid chromatography and fluorescence polarization
Steven J McCann1, Leslie O White, Brian Keevil
1Department of Clinical Biochemistry, Wythenshawe Hospital, Southmoor Road, Manchester M23 9LT, UK. steven.mccann@smuht.nwest.nhs.uk
Abstract:
A simple liquid extraction coupled with reverse-phase HPLC and UV detection was shown to correlate well with fluorescence polarization immunoassay (FPIA) on the Abbott TD(x) analyser for serum teicoplanin analysis, r(2) = 0.974, HPLC = 0.908 TDx + 2.324. A Bland-Altman plot showed no significant bias between the results. The HPLC method was linear over the range 10-100 mg/L. The HPLC method showed very good reproducibility, comparable to FPIA. The inter-assay coefficients of variation were 2.76%, 2.29% and 2.65% at 10.3, 51.4 and 77.1 mg/L, respectively, with intra-assay coefficients of variation of 1.86%, 1.97% and 1.17% at 9.13, 50.65 and 75.1 mg/L, respectively. Recoveries were between 99.1 and 101.8% within the analytical range. The HPLC method described is simple, robust, highly reproducible and suited to a clinical laboratory with the appropriate equipment.
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