Mitsui Y, Tanimori H, Kitagawa T, Fujimaki Y, Aoki Y
Department of Parasitology, Institute of Tropical Medicine, Nagasaki University, Japan.
Am J Trop Med Hyg. 1996 Mar;54(3):243-8. doi: 10.4269/ajtmh.1996.54.243.
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.
开发了一种灵敏且可重复的酶联免疫吸附测定法（ELISA），用于测定生物体液中伊维菌素（IVM）的浓度。IVM与牛血清白蛋白和聚-L-赖氨酸的缀合物分别用于在兔体内产生抗体，并作为滴定抗体的固相标记物。竞争性ELISA的操作是，将IVM和IVM-生物素缀合物与抗IVM抗血清同时在山羊抗兔IgG（Fc）上孵育，然后用抗生物素蛋白-过氧化物酶缀合物作为示踪剂测定结合的IVM-生物素的量。该测定法在0.3-10 ng/ml范围内的变异系数小于10%。检测限为0.1 ng/ml。抗IVM抗血清与某些驱虫药的交叉反应可忽略不计。使用这种ELISA，在单剂量口服500微克/千克体重后的72小时内，很容易测定长爪沙鼠（Meriones unguiculatus）血清中的IVM水平。
