Published: Journal of Analytical Toxicology, Volume 22, Number 7, November/December 1998, pp. 573-579
Validation of an Automated Microplate Enzyme Immunoassay
for Screening of Postmortem Blood for Drugs of Abuse
Vina R. Spiehler, Ines B. Collison, Paul R. Sedgwick, Susan L. Perez, Sam
D. Le, and Dawn Ann Farnin
The objective of this study was to compare the sensitivity and specificity
of an enzyme immunoassay employing antibodies bound to a microtiter plate (MPEIA)
with those of two radioimmunoassays for screening postmortem blood from selected
coroner’s cases for drugs of abuse. The radioimmunoassays were a coated-tube
radioimmunoassay (CTRIA) and a double antibody radioimmunoassay (DARIA). Specimens
consisted of 260 postmortem blood specimens from coroner’s cases. Immunoassay
results (positive or negative) were compared with confirmed results on those
cases by gas chromatography–mass spectrometry, alone or in combination
with gas–liquid chromatography using either a nitrogen-phosphorus or flame-ionization
detector. Sensitivity was calculated as the true-positive rate using chromatographic
confirmation as the reference standard. Specificity was calculated as the true-negative
rate. Sensitivity and specificity were calculated for 5–7 potential cutoff
concentrations for the drug classes opiates, amphetamines, cocaine and metabolites,
and barbiturates. For opiates, the sensitivity and specificity were 99% and
93%, respectively, for the MPEIA at a cutoff of 20-ng/mL morphine, compared
with 94% and 96% for the CTRIA at a cutoff of 5-ng/mL morphine and > 99%
and 96% for the DARIA at 20-ng/mL morphine. For cocaine and metabolites, the
sensitivity and specificity were 96% and 93%, respectively, for the MPEIA at
50-ng/mL benzoylecgonine, compared with 93% and 96% for CTRIA at 50-ng/mL benzoylecgonine
and 98% and 97% for the DARIA at 50-ng/mL benzoylecgonine. For amphetamines,
the sensitivity and specificity were > 99% and 91%, respectively, for the
MPEIA at 25-ng/mL methamphetamine, compared with 93% and 86% for the CTRIA at
25-ng/mL methamphetamine and 83% and 89% for the DARIA at 50-ng/mL methamphetamine.
For barbiturates, the sensitivity and specificity were > 99% and 92%, respectively,
for the MPEIA at 50-ng/mL secobarbital, compared with 91% and 87% for the CTRIA
at 500-ng/mL secobarbital
and 79% and 95% for the DARIA at a cutoff of 1000-ng/mL phenobarbital.
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