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Molecular Methods for the Detection of Toxoplasma gondii Oocysts in Fresh Produce: An Extensive Review
Human infection with the important zoonotic foodborne pathogen Toxoplasma gondii has been associated with unwashed raw fresh produce consumption. The lack of a standardised detection method limits the estimation of fresh produce as an infection source. To support method development and standardisati...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7828537/ https://www.ncbi.nlm.nih.gov/pubmed/33451081 http://dx.doi.org/10.3390/microorganisms9010167 |
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author | Slana, Iva Bier, Nadja Bartosova, Barbora Marucci, Gianluca Possenti, Alessia Mayer-Scholl, Anne Jokelainen, Pikka Lalle, Marco |
author_facet | Slana, Iva Bier, Nadja Bartosova, Barbora Marucci, Gianluca Possenti, Alessia Mayer-Scholl, Anne Jokelainen, Pikka Lalle, Marco |
author_sort | Slana, Iva |
collection | PubMed |
description | Human infection with the important zoonotic foodborne pathogen Toxoplasma gondii has been associated with unwashed raw fresh produce consumption. The lack of a standardised detection method limits the estimation of fresh produce as an infection source. To support method development and standardisation, an extensive literature review and a multi-attribute assessment were performed to analyse the key aspects of published methods for the detection of T. gondii oocyst contamination in fresh produce. Seventy-seven published studies were included, with 14 focusing on fresh produce. Information gathered from expert laboratories via an online questionnaire were also included. Our findings show that procedures for oocyst recovery from fresh produce mostly involved sample washing and pelleting of the washing eluate by centrifugation, although washing procedures and buffers varied. DNA extraction procedures including mechanical or thermal shocks were identified as necessary steps to break the robust oocyst wall. The most suitable DNA detection protocols rely on qPCR, mostly targeting the B1 gene or the 529 bp repetitive element. When reported, validation data for the different detection methods were not comparable and none of the methods were supported by an interlaboratory comparative study. The results of this review will pave the way for an ongoing development of a widely applicable standard operating procedure. |
format | Online Article Text |
id | pubmed-7828537 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-78285372021-01-25 Molecular Methods for the Detection of Toxoplasma gondii Oocysts in Fresh Produce: An Extensive Review Slana, Iva Bier, Nadja Bartosova, Barbora Marucci, Gianluca Possenti, Alessia Mayer-Scholl, Anne Jokelainen, Pikka Lalle, Marco Microorganisms Review Human infection with the important zoonotic foodborne pathogen Toxoplasma gondii has been associated with unwashed raw fresh produce consumption. The lack of a standardised detection method limits the estimation of fresh produce as an infection source. To support method development and standardisation, an extensive literature review and a multi-attribute assessment were performed to analyse the key aspects of published methods for the detection of T. gondii oocyst contamination in fresh produce. Seventy-seven published studies were included, with 14 focusing on fresh produce. Information gathered from expert laboratories via an online questionnaire were also included. Our findings show that procedures for oocyst recovery from fresh produce mostly involved sample washing and pelleting of the washing eluate by centrifugation, although washing procedures and buffers varied. DNA extraction procedures including mechanical or thermal shocks were identified as necessary steps to break the robust oocyst wall. The most suitable DNA detection protocols rely on qPCR, mostly targeting the B1 gene or the 529 bp repetitive element. When reported, validation data for the different detection methods were not comparable and none of the methods were supported by an interlaboratory comparative study. The results of this review will pave the way for an ongoing development of a widely applicable standard operating procedure. MDPI 2021-01-13 /pmc/articles/PMC7828537/ /pubmed/33451081 http://dx.doi.org/10.3390/microorganisms9010167 Text en © 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Review Slana, Iva Bier, Nadja Bartosova, Barbora Marucci, Gianluca Possenti, Alessia Mayer-Scholl, Anne Jokelainen, Pikka Lalle, Marco Molecular Methods for the Detection of Toxoplasma gondii Oocysts in Fresh Produce: An Extensive Review |
title | Molecular Methods for the Detection of Toxoplasma gondii Oocysts in Fresh Produce: An Extensive Review |
title_full | Molecular Methods for the Detection of Toxoplasma gondii Oocysts in Fresh Produce: An Extensive Review |
title_fullStr | Molecular Methods for the Detection of Toxoplasma gondii Oocysts in Fresh Produce: An Extensive Review |
title_full_unstemmed | Molecular Methods for the Detection of Toxoplasma gondii Oocysts in Fresh Produce: An Extensive Review |
title_short | Molecular Methods for the Detection of Toxoplasma gondii Oocysts in Fresh Produce: An Extensive Review |
title_sort | molecular methods for the detection of toxoplasma gondii oocysts in fresh produce: an extensive review |
topic | Review |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7828537/ https://www.ncbi.nlm.nih.gov/pubmed/33451081 http://dx.doi.org/10.3390/microorganisms9010167 |
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