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A Modified IF-test to Demonstrate IgM Antibodies to Babesia Divergens of Cattle

Modifierad indirekt immunofluorescensteknik för påvisande av IgM-antikroppar mot Babesia divergens infektion hos nötkreatur

Abstract

An IF–test modified to increase sensitivity was used to detect the presence of Babesia divergens specific IgM antibodies. It consisted of three steps: (1) the incubation of Babesia antigen with test serum, followed by; (2) its incubation with protein A; and (3) the addition of rabbit anti-bovine IgM bound to protein A, which was bound with FITC. Compared to the conventional IF–technique, this modified IF-technique detected bovine anti-Babesia-IgM in test serum at a two–fold lower dilution and with a diminished background staining.

This modified IF-technique improves the possibilities for studying class Μ antibody in the serum of cattle which have been infected with B. divergens. It is possible to determine whether a calf s antibody response is due to maternally transmitted antibodies or actively acquired antibodies by determining the presence of each of the classes of immunoglobulin against B. divergens present in the calf s serum.

Sammannfattning

Jämförelse av ett konventionellt utfört IF-test med ett modifierat test i vilket utnyttjas FITC-protein A konjugerat antiglobulin samt blockering av protein A receptorer på antigenet.

För att öka den indirekta immunofluorescens(IF)-teknikens känslighet och därmed kunna påvisa närvaro av IgM-antikroppar specifika mot B. divergens utfördes testet på följande sätt: 1) inkubering av Babesia-antigen med testserum, 2) följt av inkubering med protein A och därefter 3) tillsattes kanin anti-nöt IgM till vilket bundits protein A bundet med FITC.

Med den modifierade IF-tekniken kan IgM-antikroppar påvisas tidigt efter en infektion med B. divergens ty reaktionen kan avläsas vid en låg serumspädning tack vare en låg bakgrund. Genom att bestämma förekomst av immunoglobuliner av olika klasser kan man avgöra om ett antikroppssvar beror av maternalt överförda antikroppar eller är resultat av en egen, förvärvad infektion.

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Acknowledgment

I wish to express my gratitude to Dr. L. Prage for introducing me into the potential uses of protein A, to Dr. T. Morén for taking care of the animals and collecting serum samples, to Ms. K. Candelöf, Ms. M. Väisänen and Ms. E. Tidblom for technical assistance and to Ms. G. Wikne for typing the manuscript.

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Christensson, D.A. A Modified IF-test to Demonstrate IgM Antibodies to Babesia Divergens of Cattle. Acta Vet Scand 28, 361–371 (1987). https://doi.org/10.1186/BF03548604

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Keywords

  • FITC-labelled protein A
  • IF-technique
  • blocking of antigen receptors