Development of a recombinase polymerase amplification lateral flow assay for the detection of active Trypanosoma evansi infections.
BACKGROUND:Animal trypanosomosis caused by Trypanosoma evansi is known as "surra" and is a widespread neglected tropical disease affecting wild and domestic animals mainly in South America, the Middle East, North Africa and Asia. An essential necessity for T. evansi infection control is th...
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ftdoajarticles:oai:doaj.org/article:bdb16b6ff20b47cbac3dcf5bf08ea51f 2023-05-15T15:16:18+02:00 Development of a recombinase polymerase amplification lateral flow assay for the detection of active Trypanosoma evansi infections. Zeng Li Joar Esteban Pinto Torres Julie Goossens Benoit Stijlemans Yann G-J Sterckx Stefan Magez 2020-02-01T00:00:00Z https://doi.org/10.1371/journal.pntd.0008044 https://doaj.org/article/bdb16b6ff20b47cbac3dcf5bf08ea51f EN eng Public Library of Science (PLoS) https://doi.org/10.1371/journal.pntd.0008044 https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0008044 https://doaj.org/article/bdb16b6ff20b47cbac3dcf5bf08ea51f PLoS Neglected Tropical Diseases, Vol 14, Iss 2, p e0008044 (2020) Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 article 2020 ftdoajarticles https://doi.org/10.1371/journal.pntd.0008044 2022-12-31T07:48:33Z BACKGROUND:Animal trypanosomosis caused by Trypanosoma evansi is known as "surra" and is a widespread neglected tropical disease affecting wild and domestic animals mainly in South America, the Middle East, North Africa and Asia. An essential necessity for T. evansi infection control is the availability of reliable and sensitive diagnostic tools. While DNA-based PCR detection techniques meet these criteria, most of them require well-trained and experienced users as well as a laboratory environment allowing correct protocol execution. As an alternative, we developed a recombinase polymerase amplification (RPA) test for Type A T. evansi. The technology uses an isothermal nucleic acid amplification approach that is simple, fast, cost-effective and is suitable for use in minimally equipped laboratories and even field settings. METHODOLOGY/PRINCIPLE FINDINGS:An RPA assay targeting the T. evansi RoTat1.2 VSG gene was designed for the DNA-based detection of T. evansi. Comparing post-amplification visualization by agarose gel electrophoresis and a lateral flow (LF) format reveals that the latter displays a higher sensitivity. The RPA-LF assay is specific for RoTat1.2-expressing strains of T. evansi as it does not detect the genomic DNA of other trypanosomatids. Finally, experimental mouse infection trials demonstrate that the T. evansi specific RPA-LF can be employed as a test-of-cure tool. CONCLUSIONS/SIGNIFICANCE:Compared to other DNA-based parasite detection methods (such as PCR and LAMP), the T. evansi RPA-LF (TevRPA-LF) described in this paper is an interesting alternative because of its simple read-out (user-friendly), short execution time (15 minutes), experimental sensitivity of 100 fg purified genomic T. evansi DNA, and ability to be carried out at a moderate, constant temperature (39°C). Therefore, the TevRPA-LF is an interesting tool for the detection of active T. evansi infections. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic PLOS Neglected Tropical Diseases 14 2 e0008044 |
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ftdoajarticles |
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English |
topic |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
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Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 Zeng Li Joar Esteban Pinto Torres Julie Goossens Benoit Stijlemans Yann G-J Sterckx Stefan Magez Development of a recombinase polymerase amplification lateral flow assay for the detection of active Trypanosoma evansi infections. |
topic_facet |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
description |
BACKGROUND:Animal trypanosomosis caused by Trypanosoma evansi is known as "surra" and is a widespread neglected tropical disease affecting wild and domestic animals mainly in South America, the Middle East, North Africa and Asia. An essential necessity for T. evansi infection control is the availability of reliable and sensitive diagnostic tools. While DNA-based PCR detection techniques meet these criteria, most of them require well-trained and experienced users as well as a laboratory environment allowing correct protocol execution. As an alternative, we developed a recombinase polymerase amplification (RPA) test for Type A T. evansi. The technology uses an isothermal nucleic acid amplification approach that is simple, fast, cost-effective and is suitable for use in minimally equipped laboratories and even field settings. METHODOLOGY/PRINCIPLE FINDINGS:An RPA assay targeting the T. evansi RoTat1.2 VSG gene was designed for the DNA-based detection of T. evansi. Comparing post-amplification visualization by agarose gel electrophoresis and a lateral flow (LF) format reveals that the latter displays a higher sensitivity. The RPA-LF assay is specific for RoTat1.2-expressing strains of T. evansi as it does not detect the genomic DNA of other trypanosomatids. Finally, experimental mouse infection trials demonstrate that the T. evansi specific RPA-LF can be employed as a test-of-cure tool. CONCLUSIONS/SIGNIFICANCE:Compared to other DNA-based parasite detection methods (such as PCR and LAMP), the T. evansi RPA-LF (TevRPA-LF) described in this paper is an interesting alternative because of its simple read-out (user-friendly), short execution time (15 minutes), experimental sensitivity of 100 fg purified genomic T. evansi DNA, and ability to be carried out at a moderate, constant temperature (39°C). Therefore, the TevRPA-LF is an interesting tool for the detection of active T. evansi infections. |
format |
Article in Journal/Newspaper |
author |
Zeng Li Joar Esteban Pinto Torres Julie Goossens Benoit Stijlemans Yann G-J Sterckx Stefan Magez |
author_facet |
Zeng Li Joar Esteban Pinto Torres Julie Goossens Benoit Stijlemans Yann G-J Sterckx Stefan Magez |
author_sort |
Zeng Li |
title |
Development of a recombinase polymerase amplification lateral flow assay for the detection of active Trypanosoma evansi infections. |
title_short |
Development of a recombinase polymerase amplification lateral flow assay for the detection of active Trypanosoma evansi infections. |
title_full |
Development of a recombinase polymerase amplification lateral flow assay for the detection of active Trypanosoma evansi infections. |
title_fullStr |
Development of a recombinase polymerase amplification lateral flow assay for the detection of active Trypanosoma evansi infections. |
title_full_unstemmed |
Development of a recombinase polymerase amplification lateral flow assay for the detection of active Trypanosoma evansi infections. |
title_sort |
development of a recombinase polymerase amplification lateral flow assay for the detection of active trypanosoma evansi infections. |
publisher |
Public Library of Science (PLoS) |
publishDate |
2020 |
url |
https://doi.org/10.1371/journal.pntd.0008044 https://doaj.org/article/bdb16b6ff20b47cbac3dcf5bf08ea51f |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
PLoS Neglected Tropical Diseases, Vol 14, Iss 2, p e0008044 (2020) |
op_relation |
https://doi.org/10.1371/journal.pntd.0008044 https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0008044 https://doaj.org/article/bdb16b6ff20b47cbac3dcf5bf08ea51f |
op_doi |
https://doi.org/10.1371/journal.pntd.0008044 |
container_title |
PLOS Neglected Tropical Diseases |
container_volume |
14 |
container_issue |
2 |
container_start_page |
e0008044 |
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1766346595068018688 |