Serologic response of Babesia equi-infected horses as measured by complement-fixation and indirect fluorescent antibody tests

Kuttler, Kenneth; Goff, Willard Lynn; Gipson, Chester; Blackburn, Billie O.

doi:10.1016/0304-4017(88)90088-x

Cited by 21 publications

(18 citation statements)

References 6 publications

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“…The end antibody titre was successfully determined by both ELISA and a rise in antibody titre was initially observed from day 9 PI onward. This observation is in close conformity with earlier reports by Kumar et al (5), Knowles et al (10), Weiland (21), and Kuttler et al (22). The antibody titre plateau after days 66-73 PI with no further rise was recorded.…”

Section: Discussionsupporting

confidence: 93%

Immunokinetics of Theileria equi specific antibodies: a comparison in serial and single dilution ELISA antibody end titres

Kumar¹,

Malhotra²,

Nichani³

et al. 2013

Turk J Vet Anim Sci

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Section: Discussionsupporting

confidence: 93%

Immunokinetics of Theileria equi specific antibodies: a comparison in serial and single dilution ELISA antibody end titres

Kumar¹,

Malhotra²,

Nichani³

et al. 2013

Turk J Vet Anim Sci

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“…Previous studies have demonstrated antigenic differences between T. equi isolates from various regions (HEUCHERT et al, 1999). Kuttler et al (1988) obtained higher titers in IFAT test using homologous antigens. Therefore, there is an increasing need to consider differences in parasite characteristics at the molecular level in order to develop more sensitive and specific diagnostic methods.…”

Section: Discussionmentioning

confidence: 88%

“…Sodium dodecyl sulfate polyacrylamide gel electrophoresis and Western blot analysis were performed as previously described (TOWBIN et al, 1979;LAEMMLI, 1970). Nitrocellulose membranes were assayed with positive T. equi and B. caballi control sera diluted at 1:100.…”

Section: Western Blotmentioning

confidence: 99%

Production of recombinant EMA-1 protein and its application for the diagnosis of Theileria equi using an enzyme immuno assay in horses from São Paulo State, Brazil

Baldani

Hilario

Nakaghi

et al. 2011

Rev. Bras. Parasitol. Vet.

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The erythrocytic-stage surface protein, Equi Merozoite Antigen 1 (EMA-1), is a major candidate for the development of a diagnostic antigen for equine piroplasmosis. In order to establish an effective diagnostic method for practical use, the gene encoding the entire EMA-1 of Theileria equi Jaboticabal strain was cloned and expressed in Escherichia coli as a histidine-tagged protein (His6-EMA1). The expressed EMA-1 reacted with specific antibodies in Western blot and had an apparent molecular mass of 34 kDa which was largely consistent with its theoretical value. The nucleotide sequence of the EMA-1 gene of Jaboticabal strain was comparatively analyzed with other published sequences. The results indicated a high degree of homology with EMA-1 genes of all other strains isolated from various countries. The recombinant purified His6-EMA1 protein was tested in an enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies anti-T. equi in horses. The ELISA clearly differentiated T. equi-infected from Babesia caballi-infected horse sera or normal horse sera. Field serum samples collected from horses in the State of São Paulo, Southeastern Brazil, were examined for the diagnosis of T. equi infection by ELISA. Of 170 samples analyzed, 95.88% (163/170) were positive for T. equi infection. These results suggest that the His6-EMA1 protein expressed in E. coli could be a reliable immunodiagnostic antigen for ELISA test and that T. equi infection is a serious concern in the State of São Paulo, Brazil.

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“…6,7,9,11,12 This report describes a single, rapid, simple protocol for the competitive ELISA (cELISA) of equine serum designed to replace the current CF procedures for the serodiagnosis of piroplasmosis, dourine, and glanders.…”

mentioning

confidence: 99%

Procedurally Similar Competitive Immunoassay Systems for the Serodiagnosis ofBabesia Equi, Babesia Caballi, Trypanosoma Equiperdum, andBurkholderia MalleiInfection in Horses

2000

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Abstract. Procedurally similar competitive enzyme-linked immunoassay (cELISA) methods were developed for the serodiagnosis of Babesia equi and Babesia caballi (piroplasmosis), Trypanosoma equiperdum (dourine), and Burkholderia mallei (glanders) infections in horses. Apparent test specificities for the B. equi, B. caballi, T. equiperdum, and B. mallei cELISAs were 99.2%, 99.5%, 98.9%, and 98.9%, respectively. Concordances and kappa values between the complement fixation (CF) and the cELISA procedures for the serodiagnosis of B. equi, B. caballi, T. equiperdum, and B. mallei infections in experimentally exposed horses were 76% and 0.55, 89% and 0.78, 97% and 0.95, and 70% and 0.44, respectively. The cELISA method may be a technically more reproducible, objective, and convenient approach for piroplasmosis, dourine, and glanders serodiagnosis in qualifying animals for international movement and disease eradication programs than the CF systems currently in use. Use of the cELISA method also obviated the problems associated with testing hemolyzed or anticomplementary sera.Piroplasmosis, dourine, and glanders are internationally reportable, serious infectious diseases of Equidae. 1,3 The etiologic agents of these diseases are unrelated. Piroplasmosis results from Babesia equi or Babesia caballi infection, whereas dourine and glanders result from Trypanosoma equiperdum and Burkholderia mallei infections respectively. 1,3 Many nations require that Equidae presented for importation be serologically negative to B. equi, B. caballi, T. equiperdum, and B. mallei. 1,3,10 Traditionally, a panel of 4 separate complement fixation (CF) procedures has been the primary serodiagnostic tool prescribed by the Office International des Epizooties (OIE) for this purpose. 2,3 The OIE has also recognized alternative serodiagnostic methods for each etiologic agent, including separate indirect fluorescent antibody tests (IFATs) for B. equi and B. caballi, enzyme-linked immunosorbent assay (ELISA) for B. mallei, and ELISA, IFAT, or agar gel immunodiffusion procedures for T. equiperdum serodiagnosis. 4,6,7,12,13,17 In practice, the battery of 4 CF procedures has been the most convenient, rapid, single test system for accomplishing the required piroplasmosis, dourine, and glanders serodiagnostic testing. Unfortunately, the CF tests require careful continuous titration of numerous labile reagents and, for piroplasmosis, the laborious expensive preparation of antigen from infected splenectomized horses. 2,3 These CF procedures do not From the Diagnostic Bacteriology Laboratory, National Veterinary Services Laboratories, US Department of Agriculture, Animal and Health Inspection Services, Ames, IA 50010.Received for publication January 19, 1999. function with sera having anticomplementary activity, test interpretation is often subjective, test sensitivity is low relative to more modern immunoassay methods, and CF test sensitivity declines as the serologic responses of exposed animals shift from initial IgMbased reactions to those of other immu...

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Serologic response of Babesia equi-infected horses as measured by complement-fixation and indirect fluorescent antibody tests

Cited by 21 publications

References 6 publications

Immunokinetics of Theileria equi specific antibodies: a comparison in serial and single dilution ELISA antibody end titres

Immunokinetics of Theileria equi specific antibodies: a comparison in serial and single dilution ELISA antibody end titres

Production of recombinant EMA-1 protein and its application for the diagnosis of Theileria equi using an enzyme immuno assay in horses from São Paulo State, Brazil

Procedurally Similar Competitive Immunoassay Systems for the Serodiagnosis ofBabesia Equi, Babesia Caballi, Trypanosoma Equiperdum, andBurkholderia MalleiInfection in Horses

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