Illicit heroin is trafficked as a solid particulate drug, while heroin
abuse is monitored by testing urine samples for its principal metabol
ites, morphine and morphine-3-glucuronide, Two novel bacterial enzymes
were used in the development of a linked-enzyme assay for heroin and
its metabolites: heroin esterase, which converts heroin to morphine, a
nd morphine dehydrogenase, which oxidizes morphine to morphinone with
the concomitant reduction of NADP(+). A bioluminescent assay involving
heroin esterase, morphine dehydrogenase, and the bacterial luciferase
from Vibrio harveyi was developed and shown to be sensitive to 89 ng/
mL heroin and 2.0 ng/mt morphine, Excellent correlation with the resul
ts from 83 authentic samples submitted for urine drug screening at a h
ospital laboratory was obtained. The bioluminescent assay exhibited gr
eater specificity and speed than current immunological screening metho
ds, A novel format of the bioluminescent assay involving immobilized e
nzymes was sensitive to 101 ng (250 pmol) of heroin and responded well
to particulate heroin. This form of the test was sensitive enough to
respond to one or two typical particles of illicit heroin.