Enzyme-linked immunosorbent assay (ELISA) for detection of anti-Trypanosoma evansi equine antibodies

Reyna-Bello, Armando; García, Francisco; Rivera, Moisés Hinojosa; Sansó, Bruno; Aso, Pedro María

doi:10.1016/s0304-4017(98)00199-x

Cited by 33 publications

(15 citation statements)

References 14 publications

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“…Several reports have shown a very high immunological cross-reactivity between T. evansi and T. vivax (Aray et al, 1998;Reyna-Bello et al, 1998;Uzcanga et al, 2002). Since, the production of T. vivax antigens is a limiting factor (Desquesnes and Tresse, 1996) as this parasite is very difficult to propagate in experimental animal models (Gardiner, 1989), our aim has been to identify and isolate antigens from T. evansi that are responsible for this cross-reaction.…”

Section: Introductionmentioning

confidence: 99%

“…In particular PCR, which is a very sensitive technique for detecting trypanosomes (Moser et al, 1989;Masiga et al, 1992;Majiwa et al, 1994;Solano and Amsler-Delafosse, 1995;Reifenberg et al, 1997;Lefrançois et al, 1998), has limited use in routine diagnosis due to the need of specific laboratory facilities, common DNA cross-contamination, and high costs. While detection of trypanosome antigens lacks sensitivity and specificity (Desquesnes, 1996;Eisler et al, 1998;Rebeski et al, 1999), assays for detection of anti-trypanosome circulating antibodies have high measures of validity and are economical and applicable at a large scale (Reyna-Bello et al, 1998;Rebeski et al, 2000;Lejon et al, 2003).…”

Section: Introductionmentioning

confidence: 99%

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Isolation of two antigens from Trypanosoma evansi that are partially responsible for its cross-reactivity with Trypanosoma vivax

Camargo

Uzcanga

Bubis

2004

Veterinary Parasitology

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Isolation of two antigens from Trypanosoma evansi that are partially responsible for its cross-reactivity with Trypanosoma vivax

Camargo

Uzcanga

Bubis

2004

Veterinary Parasitology

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“…Nevertheless, a total of 61 parasite-positive camels, antibody-detection enzyme-linked immunosorbent assay (Ab-ELISA) detected 95% (OLAHO-MUKANI et al, 1993). Reyna-Bello et al, (1998) standardized the ELISA for detection of anti-T. evansi antibodies in naturally and experimentally infected horses. The results obtained were compared to those from an IFAT, with a relative sensibility of 98.3%, a specificity of 95.12% and a predictive value of 96.83% were determined.…”

Section: Resultsmentioning

confidence: 99%

Resposta imune humoral de eqüinos infectados experimentalmente com Trypanosoma evansi

et al. 2001

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Resumo: Seis eqüinos foram inoculados com 10 6 tripomastigota sangüícolas de Trypanosoma evansi. Três outros animais foram mantidos como testemunhas. Amostras de soro sangüíneo foram obtidas de todos os animais, antes da inoculação, e diariamente até o 14º dia pós inoculação (DPI); após este período uma vez por semana. Pesquisa de anticorpos anti-T. evansi, foram realizadas através da reação de imunofluorescência indireta (RIFI) e do ensaio de imunoabsorção enzimática (ELISA). A resposta imune humoral, detectada através da RIFI e do ELISA, iniciou-se, em média, a partir do oitavo DPI, alcançando títulos máximos após quatro semanas de evolução, e os titulos de anticorpos anti-T. evansi mantiveram-se elevadas até o término das observações. Palavras-chave: Trypanosoma evansi, tripanossomiase, eqüinos, resposta imune.Abstract: Six adult horses were experimentally infected with Trypanosoma evansi (10 6 parasites). Three other adult horses served as negative control. Serum samples of the experimentally infected horses with T. evansi and non-infected controls horses were obtained before inoculation, and daily thereafter until 14 days post infection (DPI). After that time the serum samples were obtained weekly. Sera of the infected and non-infected control horses was tested by indirect fluorescent antibody test (IFAT) and enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies against T. evansi. Both ELISA and IFAT detected trypanosomal antibodies shortly after infection and showed progressive increases in antibodies levels during early stages of infection. The responses started on the eighth and eleventh DPI. Maximum IFAT and ELISA values were reached after four weeks of infection and were maintained at this level until the end of the period of study.

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“…Due to the cross reaction between some trypanosomes, [1,12] and specifically between T. evansi and T. vivax, [9] the ELISA was performed in order to detect bovine anti-T. vivax using T. evansi as the antigen. The soluble extract was purified according to Reyna-Bello et al [9] Briefly, a cryopreserved sample of T. evansi was inoculated in a rat (Spragüe Dawley) and parasitemia was followed until it reached 10 9 parasites/mL.…”

Section: Soluble Antigenic Extract From T Evansimentioning

confidence: 99%

“…The soluble extract was purified according to Reyna-Bello et al [9] Briefly, a cryopreserved sample of T. evansi was inoculated in a rat (Spragüe Dawley) and parasitemia was followed until it reached 10 9 parasites/mL. The parasites were purified using a DEAE-Cellulose column, [13] and then centrifuged at 16000 Â g. The precipitate was resuspended in PBS at pH 8, and sonicated in five The 96 polyvinyle wells (Nunc, Polysop, Denmark) were sensitized 100 mL/ well with the antigenic extract isolated from T. evansi, diluted in 50 mM of carbonate bicarbonate buffer at pH 9.6 at a concentration of 32 and 8 mg/mL.…”

Section: Soluble Antigenic Extract From T Evansimentioning

confidence: 99%

Assessment of Chromogen Suitability in ELISA for the Detection of Anaplasmosis and Trypanosomosis

Reyna-Bello

Eleizalde

Silva

2007

Journal of Immunoassay and Immunochemistry

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Two different ELISAs were routinely performed in our laboratory to detect bovine trypanosomosis and anaplasmosis. The ELISA test for trypanosomosis involved the adsorption of a soluble fraction of parasites as the antigen; and, the ELISA for anaplasmosis was performed with a purified recombinant protein MSP5r adsorbed to the plate. With the purpose of assessing the merit of ABTS and TMB, we compared the absorbance obtained from positive and negative control sera from both assays. The results obtained, suggest that TMB is more adequate for recombinant antigens and that ABTS is preferred when partially purified antigenic extracts are used in the ELISA test.

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Enzyme-linked immunosorbent assay (ELISA) for detection of anti-Trypanosoma evansi equine antibodies

Cited by 33 publications

References 14 publications

Isolation of two antigens from Trypanosoma evansi that are partially responsible for its cross-reactivity with Trypanosoma vivax

Isolation of two antigens from Trypanosoma evansi that are partially responsible for its cross-reactivity with Trypanosoma vivax

Resposta imune humoral de eqüinos infectados experimentalmente com Trypanosoma evansi

Assessment of Chromogen Suitability in ELISA for the Detection of Anaplasmosis and Trypanosomosis

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