A competitive enzyme-linked immunosorbent assay for diminazene.

Wycliff M Karanja, Raymond E Mdachi, Grace A Murilla
Author Information
  1. Wycliff M Karanja: Kenya Trypanosomiasis Research Institute (KETRI), PO Box 362, Kikuyu, Kenya. ketri@net2000ke.com

Abstract

diminazene aceturate has remained a very important therapeutic drug for trypanosomosis in cattle, sheep and goats since its introduction into the market in 1955. Despite its continued use, the methods available for its detection in body fluids are lengthy and inefficient for routine monitoring of drug levels in treated animals. A competitive enzyme linked immunosorbent assay (ELISA) has now been developed and optimized for the detection of diminazene in bovine serum. In the assay, diminazene in the test samples and that in a newly developed diminazene-horseradish peroxidase conjugate compete for antibodies to diminazene raised in rabbits and immobilized on a microtitre plate. Tetramethylbenzidine-hydrogen peroxide (TMB/H(2)O(2)) is used as chromogen-substrate system. The assay has a detection limit of 0.8 ng/ml of serum with a high specificity for diminazene. Cross-reactivity with either homidium bromide and quinapyramine sulphate/chloride of 0.0004% is negligible while that with isometamidium chloride is 0.71%. The assay was able to detect diminazene levels in normal Boran steers for at least two weeks after intramuscular injection with the drug at a dose of 3.5 mg/kg bw. The assay will be useful in monitoring diminazene use, and development of resistance in trypanosomosis endemic areas.

MeSH Term

Animals
Cattle
Diminazene
Enzyme-Linked Immunosorbent Assay
Half-Life
Rabbits
Trypanocidal Agents

Chemicals

Trypanocidal Agents
Diminazene

Word Cloud

Created with Highcharts 10.0.0diminazeneassaydrugdetection0trypanosomosisusemonitoringlevelscompetitiveimmunosorbentdevelopedserum2Diminazeneaceturateremainedimportanttherapeuticcattlesheepgoatssinceintroductionmarket1955DespitecontinuedmethodsavailablebodyfluidslengthyinefficientroutinetreatedanimalsenzymelinkedELISAnowoptimizedbovinetestsamplesnewlydiminazene-horseradishperoxidaseconjugatecompeteantibodiesraisedrabbitsimmobilizedmicrotitreplateTetramethylbenzidine-hydrogenperoxideTMB/HOusedchromogen-substratesystemlimit8ng/mlhighspecificityCross-reactivityeitherhomidiumbromidequinapyraminesulphate/chloride0004%negligibleisometamidiumchloride71%abledetectnormalBoransteersleasttwoweeksintramuscularinjectiondose35mg/kgbwwillusefuldevelopmentresistanceendemicareasenzyme-linked

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