Comparison of an ELISA assay for the detection of adhesive/invasive Neospora caninumtachyzoites

Neospora caninum belongs to the phylum Apicomplexa, the causative agent of neosporosis, which leads to economic impacts on cattle production. A common feature among apicomplexan parasites is the invasive process driven mostly by the parasite. As a first evaluation of candidate molecules that play a possible role by interfering in this invasive process, the in vitro invasion assay is a fast and direct way to screen future agonists or antagonists. This work involved the development of a new cell culture ELISA and transient β-galactosidase activity applied to the semi-quantitative detection of N. caninum in Vero cell culture. Cell culture ELISA is based on histochemistry and immunology, resulting in a colorimetric reaction. The β-galactosidase activity was obtained by the transient transfection of the lacZ gene under control of RPS13 promoter of N. caninum. These methods were used to evaluate the effects of temperature (37°C and 85°C) on the invasion and adhesion of tachyzoites. The three tested methods (real time PCR, β-galactosidase activity and ELISA) showed a similar pattern, indicating that different methods may be complementary.

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Main Authors: Pereira,Luiz Miguel, Yatsuda,Ana Patrícia
Format: Digital revista
Language:English
Published: Colégio Brasileiro de Parasitologia Veterinária 2014
Online Access:http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1984-29612014000100006
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spelling oai:scielo:S1984-296120140001000062015-09-23Comparison of an ELISA assay for the detection of adhesive/invasive Neospora caninumtachyzoitesPereira,Luiz MiguelYatsuda,Ana Patrícia Neospora caninum cell culture ELISA LacZ gene transfection real-time PCR invasion assay Neospora caninum belongs to the phylum Apicomplexa, the causative agent of neosporosis, which leads to economic impacts on cattle production. A common feature among apicomplexan parasites is the invasive process driven mostly by the parasite. As a first evaluation of candidate molecules that play a possible role by interfering in this invasive process, the in vitro invasion assay is a fast and direct way to screen future agonists or antagonists. This work involved the development of a new cell culture ELISA and transient β-galactosidase activity applied to the semi-quantitative detection of N. caninum in Vero cell culture. Cell culture ELISA is based on histochemistry and immunology, resulting in a colorimetric reaction. The β-galactosidase activity was obtained by the transient transfection of the lacZ gene under control of RPS13 promoter of N. caninum. These methods were used to evaluate the effects of temperature (37°C and 85°C) on the invasion and adhesion of tachyzoites. The three tested methods (real time PCR, β-galactosidase activity and ELISA) showed a similar pattern, indicating that different methods may be complementary.info:eu-repo/semantics/openAccessColégio Brasileiro de Parasitologia VeterináriaRevista Brasileira de Parasitologia Veterinária v.23 n.1 20142014-03-01info:eu-repo/semantics/articletext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S1984-29612014000100006en10.1590/S1984-29612014005
institution SCIELO
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country Brasil
countrycode BR
component Revista
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region America del Sur
libraryname SciELO
language English
format Digital
author Pereira,Luiz Miguel
Yatsuda,Ana Patrícia
spellingShingle Pereira,Luiz Miguel
Yatsuda,Ana Patrícia
Comparison of an ELISA assay for the detection of adhesive/invasive Neospora caninumtachyzoites
author_facet Pereira,Luiz Miguel
Yatsuda,Ana Patrícia
author_sort Pereira,Luiz Miguel
title Comparison of an ELISA assay for the detection of adhesive/invasive Neospora caninumtachyzoites
title_short Comparison of an ELISA assay for the detection of adhesive/invasive Neospora caninumtachyzoites
title_full Comparison of an ELISA assay for the detection of adhesive/invasive Neospora caninumtachyzoites
title_fullStr Comparison of an ELISA assay for the detection of adhesive/invasive Neospora caninumtachyzoites
title_full_unstemmed Comparison of an ELISA assay for the detection of adhesive/invasive Neospora caninumtachyzoites
title_sort comparison of an elisa assay for the detection of adhesive/invasive neospora caninumtachyzoites
description Neospora caninum belongs to the phylum Apicomplexa, the causative agent of neosporosis, which leads to economic impacts on cattle production. A common feature among apicomplexan parasites is the invasive process driven mostly by the parasite. As a first evaluation of candidate molecules that play a possible role by interfering in this invasive process, the in vitro invasion assay is a fast and direct way to screen future agonists or antagonists. This work involved the development of a new cell culture ELISA and transient β-galactosidase activity applied to the semi-quantitative detection of N. caninum in Vero cell culture. Cell culture ELISA is based on histochemistry and immunology, resulting in a colorimetric reaction. The β-galactosidase activity was obtained by the transient transfection of the lacZ gene under control of RPS13 promoter of N. caninum. These methods were used to evaluate the effects of temperature (37°C and 85°C) on the invasion and adhesion of tachyzoites. The three tested methods (real time PCR, β-galactosidase activity and ELISA) showed a similar pattern, indicating that different methods may be complementary.
publisher Colégio Brasileiro de Parasitologia Veterinária
publishDate 2014
url http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1984-29612014000100006
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