Analytical sensitivity of a multiplex quantitative PCR for Toxoplasma gondii and Neospora caninum

Cyst-forming coccidia, Toxoplasma gondii and Neospora caninum, are recognised as important causes of animal disease. Molecular diagnostics based on the presence of DNA in animal tissue are required to specifically detect T. gondii and N. caninum while achieving high levels of analytical sensitivity....

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Autores principales: Truong, Marcus, Šlapeta, Jan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Berlin Heidelberg 2023
Materias:
Brief Report
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10006017/
https://www.ncbi.nlm.nih.gov/pubmed/36806974
http://dx.doi.org/10.1007/s00436-023-07796-5
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author Truong, Marcus
Šlapeta, Jan
author_facet Truong, Marcus
Šlapeta, Jan
author_sort Truong, Marcus
collection PubMed
description Cyst-forming coccidia, Toxoplasma gondii and Neospora caninum, are recognised as important causes of animal disease. Molecular diagnostics based on the presence of DNA in animal tissue are required to specifically detect T. gondii and N. caninum while achieving high levels of analytical sensitivity. We optimised available single-plex probe base qPCR assays into a multiplexed qPCR panel to detect cyst-forming coccidia, i.e. T. gondii and N. caninum. The T. gondii assay is based on a 529-bp repetitive (REP) element and the N. caninum assay on the NC5 repetitive region. Using target sequence synthetic DNA, the limit of detection (LOD) was determined to be 100 copies, that is less than a single tachyzoite of either T. gondii or N. caninum. The T. gondii and N. caninum multiplexed qPCR assay optimised in this study can be used to effectively detect parasite DNA for diagnostic purposes in animal tissue.
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spelling pubmed-100060172023-03-12 Analytical sensitivity of a multiplex quantitative PCR for Toxoplasma gondii and Neospora caninum Truong, Marcus Šlapeta, Jan Parasitol Res Brief Report Cyst-forming coccidia, Toxoplasma gondii and Neospora caninum, are recognised as important causes of animal disease. Molecular diagnostics based on the presence of DNA in animal tissue are required to specifically detect T. gondii and N. caninum while achieving high levels of analytical sensitivity. We optimised available single-plex probe base qPCR assays into a multiplexed qPCR panel to detect cyst-forming coccidia, i.e. T. gondii and N. caninum. The T. gondii assay is based on a 529-bp repetitive (REP) element and the N. caninum assay on the NC5 repetitive region. Using target sequence synthetic DNA, the limit of detection (LOD) was determined to be 100 copies, that is less than a single tachyzoite of either T. gondii or N. caninum. The T. gondii and N. caninum multiplexed qPCR assay optimised in this study can be used to effectively detect parasite DNA for diagnostic purposes in animal tissue. Springer Berlin Heidelberg 2023-02-18 2023 /pmc/articles/PMC10006017/ /pubmed/36806974 http://dx.doi.org/10.1007/s00436-023-07796-5 Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Brief Report
Truong, Marcus
Šlapeta, Jan
Analytical sensitivity of a multiplex quantitative PCR for Toxoplasma gondii and Neospora caninum
title Analytical sensitivity of a multiplex quantitative PCR for Toxoplasma gondii and Neospora caninum
title_full Analytical sensitivity of a multiplex quantitative PCR for Toxoplasma gondii and Neospora caninum
title_fullStr Analytical sensitivity of a multiplex quantitative PCR for Toxoplasma gondii and Neospora caninum
title_full_unstemmed Analytical sensitivity of a multiplex quantitative PCR for Toxoplasma gondii and Neospora caninum
title_short Analytical sensitivity of a multiplex quantitative PCR for Toxoplasma gondii and Neospora caninum
title_sort analytical sensitivity of a multiplex quantitative pcr for toxoplasma gondii and neospora caninum
topic Brief Report
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10006017/
https://www.ncbi.nlm.nih.gov/pubmed/36806974
http://dx.doi.org/10.1007/s00436-023-07796-5
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