Molecular Methods for the Detection of Toxoplasma gondii Oocysts in Fresh Produce: An Extensive Review

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00027162%3A_____%2F21%3AN0000006" target="_blank" >RIV/00027162:_____/21:N0000006 - isvavai.cz</a>

Výsledek na webu

<a href="https://www.mdpi.com/2076-2607/9/1/167" target="_blank" >https://www.mdpi.com/2076-2607/9/1/167</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.3390/microorganisms9010167" target="_blank" >10.3390/microorganisms9010167</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Molecular Methods for the Detection of Toxoplasma gondii Oocysts in Fresh Produce: An Extensive Review

Popis výsledku v původním jazyce

Toxoplasma gondii is an important zoonotic foodborne pathogen worldwide and infection in humans has been associated with the consumption of unwashed raw fresh produce. The estimation of relative importance of fresh produce as an infection source is however hampered by the lack of a standardised detection method Aim To support method development and standardisation, we aimed to summarize the key aspects of published methods for the detection of T. gondii oocyst contamination in fresh produce. Methods We performed an extensive literature review and a multi-attribute assessment of published T. gondii oocyst detection methods, irrespective of the contamination matrix, by focusing on three steps: oocyst recovery, DNA extraction, and DNA amplification. The literature review was further supplemented with input from a questionnaire study administered online to 24 expert laboratories worldwide. Results Seventy-seven published studies were included, with 14 focusing on fresh produce. 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 step to break the robust oocyst wall. The most suitable DNA detection protocols rely on qPCR, mostly targeting the B1 gene or the 5 29 bp repetitive element. When reported, validation data for the different detection methods were not comparable and none of the methods was supported by an interlaboratory comparative study. Conclusions The key step to enable detection is oocyst recovery, followed by an efficient DNA extraction and sensitive and specific amplification of the parasite’s DNA. The results of this review will pave the way for an ongoing development of a widely applicable standard operating procedure.

Název v anglickém jazyce

Molecular Methods for the Detection of Toxoplasma gondii Oocysts in Fresh Produce: An Extensive Review

Popis výsledku anglicky

Toxoplasma gondii is an important zoonotic foodborne pathogen worldwide and infection in humans has been associated with the consumption of unwashed raw fresh produce. The estimation of relative importance of fresh produce as an infection source is however hampered by the lack of a standardised detection method Aim To support method development and standardisation, we aimed to summarize the key aspects of published methods for the detection of T. gondii oocyst contamination in fresh produce. Methods We performed an extensive literature review and a multi-attribute assessment of published T. gondii oocyst detection methods, irrespective of the contamination matrix, by focusing on three steps: oocyst recovery, DNA extraction, and DNA amplification. The literature review was further supplemented with input from a questionnaire study administered online to 24 expert laboratories worldwide. Results Seventy-seven published studies were included, with 14 focusing on fresh produce. 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 step to break the robust oocyst wall. The most suitable DNA detection protocols rely on qPCR, mostly targeting the B1 gene or the 5 29 bp repetitive element. When reported, validation data for the different detection methods were not comparable and none of the methods was supported by an interlaboratory comparative study. Conclusions The key step to enable detection is oocyst recovery, followed by an efficient DNA extraction and sensitive and specific amplification of the parasite’s DNA. The results of this review will pave the way for an ongoing development of a widely applicable standard operating procedure.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

30310 - Parasitology

Projekt

—

Návaznosti

R - Projekt Ramcoveho programu EK

Rok uplatnění

2021

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Microorganisms

ISSN

2076-2607

e-ISSN

2076-2607

Svazek periodika

9

Číslo periodika v rámci svazku

1

Stát vydavatele periodika

CH - Švýcarská konfederace

Počet stran výsledku

19

Strana od-do

"167"

Kód UT WoS článku

000610595900001

EID výsledku v databázi Scopus

2-s2.0-85099562587