Development and analytic validation of a sandwich ELISA for the measurement of α1-proteinase inhibitor concentrations in serum and feces of common marmosets (Callithrix jacchus).
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OBJECTIVE To develop and validate a sandwich ELISA for the measurement of α1-proteinase inhibitor (α1-PI) concentrations in serum and fecal samples obtained from common marmosets (Callithrix jacchus). SAMPLE Leftover serum (n = 42) and fecal (23) samples submitted for diagnostic testing; paired serum and fecal samples obtained from 30 common marmosets at 2 research colonies. PROCEDURES A sandwich ELISA was developed and analytically validated by determining the lower limit of detection, linearity, accuracy, precision, and reproducibility. Reference intervals for α1-PI concentrations in serum and feces of common marmosets were calculated. RESULTS The standard curve was generated for concentrations between 1 and 100 ng/mL. Mean ± SD observed-to-expected ratio for serial dilutions of serum and fecal samples was 117.1 ± 5.6% (range, 112.2% to 123.0%) and 106.1 ± 19.7% (range, 82.6% to 130.2%), respectively. Mean observed-to-expected ratio for spiking recovery of serum and fecal samples was 102.9 ± 12.1% (range, 86.8% to 115.8%) and 97.9 ± 19.0% (range, 83.0% to 125.1%), respectively. Reference interval for serum concentrations of α1-PI was 1,254 to 1,813 μg/mL, for 3-day mean fecal concentrations was 11.5 to 42.2 μg/g of feces, and for 3-day maximum fecal concentrations was 13.2 to 51.2 μg/g of feces. CONCLUSIONS AND CLINICAL RELEVANCE The ELISA was linear, accurate, precise, and reproducible for quantification of α1-PI concentrations in serum and feces of common marmosets. However, the ELISA had limited linearity and accuracy for spiking recovery of fecal samples.
author list (cited authors)
Parambeth, J. C., Lidbury, J. A., Suchodolski, J. S., & Steiner, J. M.
complete list of authors
Parambeth, Joseph C||Lidbury, Jonathan A||Suchodolski, Jan S||Steiner, Jörg M