Development of a competitive enzyme-linked immunosorbent assay for diethylcarbamazine.

Y Mitsui, N Takamura, Y Fujimaki, T Yamaguchi, T Kitagawa, Y Aoki
Author Information
  1. Y Mitsui: Institute of Tropical Medicine, Nagasaki University, Japan.

Abstract

A sensitive and reproducible competitive enzyme-linked immunosorbent assay (ELISA) for the determination of the concentration of diethylcarbamazine (DEC) in biological fluids was developed. Since DEC has no functional group to conjugate with bovine serum albumin (BSA), N-(2-aminoethyl)-N-ethyl-4-methyl-1-piperazinecarboxamide (DEC-NH2) was first synthesized. This compound was then converted to carboxyl DEC (DEC-COOH) and conjugated to BSA and to poly-L-lysine for use as immunogen and solid-phase marker, respectively. The competitive ELISA was conducted by simultaneously incubating DEC with mouse anti-DEC antiserum over DEC-poly-L-lysine solid phase. Subsequently, the binding of anti-DEC antibody was detected by using sheep anti-mouse IgG peroxidase conjugate as a tracer. The reliability, determined by the coefficient of variation for inter and intra-assay, was satisfactory. The cross-reactivities of anti-DEC antibodies with DEC metabolites, related compounds and ivermectin were negligible. Using this assay, DEC levels were easily determined in serum of Mongolian jirds (Meriones unguiculatus) up to 4 hours following a single dose of DEC citrate base (100 mg/kg of body weight) via intraperitoneal route.

MeSH Term

Animals
Antibodies
Diethylcarbamazine
Enzyme-Linked Immunosorbent Assay
Female
Gerbillinae
Humans
Immune Sera
Mice

Chemicals

Antibodies
Immune Sera
Diethylcarbamazine

Word Cloud

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