Simple and sensitive enzyme-linked immunosorbent assay for ivermectin.
A sensitive and reproducible enzyme-linked immunosorbent assay (ELISA) for the determination of the concentration of ivermectin (IVM) in biological fluids was developed. A conjugate of IVM on bovine serum albumin and poly-L-lysine was used to produce antibodies in rabbits and served as a solid-phase marker for titration of antibodies, respectively. The competitive ELISA was conducted by simultaneously incubating IVM and IVM-biotin conjugate with anti-IVM antiserum over goat anti-rabbit IgG (Fc) and then determining the amount of bound IVM-biotin with avidin-peroxidase conjugate as a tracer. The coefficient of variation for the assay was less than 10% in the range of 0.3-10 ng/ml. The limit of detection was 0.1 ng/ml. The cross-reactivities of anti-IVM antiserum with some anthelmintic drugs were negligible. Using this ELISA, serum levels of IVM were easily determined in Mongolian jirds (Meriones unguiculatus) up to 72 hr following a single oral dose of 500 microgram/kg of body weight.
Mitsui
Y
Y
Department of Parasitology, Institute of Tropical Medicine, Nagasaki University, Japan.
Tanimori
H
H
Kitagawa
T
T
Fujimaki
Y
Y
Aoki
Y
Y
D000818:Animals; D000917:Antibody Formation; D000977:Antiparasitic Agents / Q000032:analysis* / Q000276:immunology / Q000493:pharmacokinetics; D001360:Avidin; D001667:Binding, Competitive; D001710:Biotin; D003429:Cross Reactions; D004306:Dose-Response Relationship, Immunologic; D004797:Enzyme-Linked Immunosorbent Assay; D005849:Gerbillinae; D007074:Immunoglobulin G; D007559:Ivermectin / Q000032:analysis* / Q000276:immunology / Q000493:pharmacokinetics; D008297:Male; D009195:Peroxidase; D011817:Rabbits; D012680:Sensitivity and Specificity; D012710:Serum Albumin, Bovine
D000977:Antiparasitic Agents; D007074:Immunoglobulin G; D001360:Avidin; D012710:Serum Albumin, Bovine; D001710:Biotin; D007559:Ivermectin; D009195:Peroxidase