Drug screening by enzyme immunoassay with the American Monitor KDA.

Abstract The methods for barbiturate, opiate, methadone, and amphetamine have been modified for use with the American Monitor KDA. The modification, which incorporates automatic correction for endogenous lysozyme activity, was evaluated by comparing results obtained with the KDA for human urine samples containing known amounts of drug(s) with results obtained with the procedure recommended by Syva for the Gilford 3500. There was 98% agreement bewteen the two systems. Six calibrators and 40 samples can be assayed for all four drugs in about 2.5 h. The procedure has proven to be reliable for screening urine samples obtained from clients at a methadone treatment center.

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Analytical and physiological validation of an enzyme immunoassay to measure oxytocin in dog, wolf, and human urine samples

Scientific Reports ◽

10.1038/s41598-021-92356-z ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

G. Wirobski ◽

F. S. Schaebs ◽

F. Range ◽

S. Marshall-Pescini ◽

T. Deschner

Keyword(s):

Enzyme Immunoassay ◽

Human Urine ◽

Extraction Efficiency ◽

Urine Samples ◽

Antibody Specificity ◽

Analytical Validation ◽

Human Urine Samples ◽

Assay Performance ◽

Fit For Purpose ◽

Commercial Enzyme Immunoassay

AbstractOxytocin (OT) promotes pro-sociality, bonding, and cooperation in a variety of species. Measuring oxytocin metabolite (OTM) concentrations in urine or saliva provides intriguing opportunities to study human and animal behaviour with minimal disturbance. However, a thorough validation of analytical methods and an assessment of the physiological significance of these measures are essential. We conducted an analytical validation of a commercial Enzyme Immunoassay (EIA; Arbor OT assay kit) to measure OTM concentrations in dog, wolf, and human urine samples. To test the assay’s ability to detect changes in OTM concentrations, we administered oxytocin intranasally to 14 dogs. Assay performance with regard to parallelism was acceptable. Assay accuracy and extraction efficiency for dog and wolf samples were comparable to a previously validated assay (Enzo OT assay kit) but variation was smaller for human samples. Binding sensitivity and antibody specificity were better in the Arbor assay. Average OTM concentrations were more than twice as high as in comparable samples measured with the Enzo assay, highlighting a lack of comparability of absolute values between different assays. Changes in OTM concentrations after intranasal treatment were detected reliably. The Arbor assay met requirements of a “fit-for-purpose” validation with improvement of several parameters compared to the Enzo assay.

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Competitive enzyme immunoassay with monoclonal antibody for homovanillic acid measurement in human urine samples

Clinical Chemistry ◽

10.1093/clinchem/43.2.363 ◽

1997 ◽

Vol 43 (2) ◽

pp. 363-368 ◽

Cited By ~ 9

Author(s):

Frédéric Taran ◽

Yveline Frobert ◽

Christophe Créminon ◽

Jacques Grassi ◽

Didier Olichon ◽

...

Keyword(s):

Monoclonal Antibody ◽

Enzyme Immunoassay ◽

Human Urine ◽

Homovanillic Acid ◽

Colorimetric Assay ◽

Cross Reactivity ◽

Urine Samples ◽

Vanillylmandelic Acid ◽

Human Urine Samples ◽

Enzyme Detection

Abstract A fast competitive enzyme immunoassay (EIA) for measuring homovanillic acid in human urine samples was developed with a monoclonal antibody and acetylcholinesterase as enzyme label. Enzyme detection was performed by an easy colorimetric assay. Monoclonal antibodies were screened on the basis of sensitivity, specificity, and correlation studies. EIA has a detection limit of 0.5 μmol/L, a CV <10% in the 1.25–10 μmol/L range, and intra- and interassay CVs of <10%. Cross-reactivity with vanillylmandelic acid was 0.5% and <8% for other structurally related catecholamine metabolites. Parallelism of the EIA was shown in dilution studies and the correlation with routine HPLC assay in 62 normal and pathologic samples was EIA = 1.492 (HPLC) − 3.46, Sy|x = 47.52, range = 4–1800 μmol/L, r2 = 0.977. Additional data concerning the validity of this assay were provided by HPLC analysis of urinary immunoreactive material.

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