Molecular Methods for the Detection of <i>Toxoplasma gondii</i> Oocysts in Fresh Produce: An Extensive Review
Human infection with the important zoonotic foodborne pathogen <i>Toxoplasma gondii</i> 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 developmen...
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| Format: | Article |
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MDPI AG
2021-01-01
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| Series: | Microorganisms |
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| Online Access: | https://www.mdpi.com/2076-2607/9/1/167 |
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| author | Iva Slana Nadja Bier Barbora Bartosova Gianluca Marucci Alessia Possenti Anne Mayer-Scholl Pikka Jokelainen Marco Lalle |
| author_facet | Iva Slana Nadja Bier Barbora Bartosova Gianluca Marucci Alessia Possenti Anne Mayer-Scholl Pikka Jokelainen Marco Lalle |
| author_sort | Iva Slana |
| collection | DOAJ |
| description | Human infection with the important zoonotic foodborne pathogen <i>Toxoplasma gondii</i> 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 <i>T. gondii</i> 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 <i>B1</i> gene or the <i>529</i> 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. |
| first_indexed | 2024-03-09T04:54:27Z |
| format | Article |
| id | doaj.art-c25654a4e9d34978b833692337374849 |
| institution | Directory Open Access Journal |
| issn | 2076-2607 |
| language | English |
| last_indexed | 2024-03-09T04:54:27Z |
| publishDate | 2021-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Microorganisms |
| spelling | doaj.art-c25654a4e9d34978b8336923373748492023-12-03T13:07:56ZengMDPI AGMicroorganisms2076-26072021-01-019116710.3390/microorganisms9010167Molecular Methods for the Detection of <i>Toxoplasma gondii</i> Oocysts in Fresh Produce: An Extensive ReviewIva Slana0Nadja Bier1Barbora Bartosova2Gianluca Marucci3Alessia Possenti4Anne Mayer-Scholl5Pikka Jokelainen6Marco Lalle7Department of Microbiology and Antimicrobial Resistance, Veterinary Research Institute, Hudcova 296/70, 621 00 Brno, Czech RepublicDepartment of Biological Safety, German Federal Institute for Risk Assessment (BfR), Max-Dohrn-Str. 8-10, 10589 Berlin, GermanyDepartment of Microbiology and Antimicrobial Resistance, Veterinary Research Institute, Hudcova 296/70, 621 00 Brno, Czech RepublicUnit of Foodborne and Neglected Parasitic Diseases, European Union Reference Laboratory for Parasites, Department of Infectious Diseases, Istituto Superiore di Sanità, viale Regina Elena 299, 00161 Rome, ItalyUnit of Foodborne and Neglected Parasitic Diseases, European Union Reference Laboratory for Parasites, Department of Infectious Diseases, Istituto Superiore di Sanità, viale Regina Elena 299, 00161 Rome, ItalyDepartment of Biological Safety, German Federal Institute for Risk Assessment (BfR), Max-Dohrn-Str. 8-10, 10589 Berlin, GermanyLaboratory of Parasitology, Infectious Disease Preparedness, Department of Bacteria, Parasites & Fungi, Statens Serum Institut, Artillerivej 5, DK-2300 Copenhagen S, DenmarkUnit of Foodborne and Neglected Parasitic Diseases, European Union Reference Laboratory for Parasites, Department of Infectious Diseases, Istituto Superiore di Sanità, viale Regina Elena 299, 00161 Rome, ItalyHuman infection with the important zoonotic foodborne pathogen <i>Toxoplasma gondii</i> 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 <i>T. gondii</i> 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 <i>B1</i> gene or the <i>529</i> 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.https://www.mdpi.com/2076-2607/9/1/167<i>Toxoplasma gondii</i>oocystready-to-eat (RTE) saladfresh producedetectiontoxoplasmosis |
| spellingShingle | Iva Slana Nadja Bier Barbora Bartosova Gianluca Marucci Alessia Possenti Anne Mayer-Scholl Pikka Jokelainen Marco Lalle Molecular Methods for the Detection of <i>Toxoplasma gondii</i> Oocysts in Fresh Produce: An Extensive Review Microorganisms <i>Toxoplasma gondii</i> oocyst ready-to-eat (RTE) salad fresh produce detection toxoplasmosis |
| title | Molecular Methods for the Detection of <i>Toxoplasma gondii</i> Oocysts in Fresh Produce: An Extensive Review |
| title_full | Molecular Methods for the Detection of <i>Toxoplasma gondii</i> Oocysts in Fresh Produce: An Extensive Review |
| title_fullStr | Molecular Methods for the Detection of <i>Toxoplasma gondii</i> Oocysts in Fresh Produce: An Extensive Review |
| title_full_unstemmed | Molecular Methods for the Detection of <i>Toxoplasma gondii</i> Oocysts in Fresh Produce: An Extensive Review |
| title_short | Molecular Methods for the Detection of <i>Toxoplasma gondii</i> Oocysts in Fresh Produce: An Extensive Review |
| title_sort | molecular methods for the detection of i toxoplasma gondii i oocysts in fresh produce an extensive review |
| topic | <i>Toxoplasma gondii</i> oocyst ready-to-eat (RTE) salad fresh produce detection toxoplasmosis |
| url | https://www.mdpi.com/2076-2607/9/1/167 |
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