A specific ELISA method for determining chloroquine in urine or dried blood spots.
Reported is an enzyme-linked immunosorbent assay (ELISA) that has been optimized and validated for the determination of chloroquine in urine or dried blood spots. The assay employs antisera raised in sheep to a chloroquine derivative conjugated to keyhole limpet haemocyanin and chloroquine conjugated to porcine thyroglobulin adsorbed onto the wells of a microtitration plate. The competitive binding of the antiserum to the wells was monitored using an alkaline-phosphatase-conjugated second antibody and a specific substrate. The assay exhibits no cross-reactivity with known chloroquine metabolites, other antimalarials, and commonly used drugs. The method was used to determine chloroquine in dried blood spot extracts and urine from a patient who was receiving a prescribed prophylactic chloroquine regimen. The drug was detected in the urine for 17 weeks and in the dried blood spots for 4 weeks after termination of the therapy.
Rowell
V
V
Rowell
F J
FJ
Baker
A
A
Laurie
D
D
Sidki
A M
AM
D000328:Adult; D000906:Antibodies / Q000032:analysis; D002738:Chloroquine / Q000097:blood / Q000276:immunology / Q000652:urine*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006801:Humans; D007106:Immune Sera / Q000302:isolation & purification; D008297:Male
D000906:Antibodies; D007106:Immune Sera; D002738:Chloroquine