Development of a droplet digital polymerase chain reaction tool for the detection of Toxoplasma gondii in meat samples.
Droplet digital polymerase chain reaction (ddPCR)
Quantitative PCR
Toxoplasma gondii
Toxoplasmosis
Journal
Parasitology research
ISSN: 1432-1955
Titre abrégé: Parasitol Res
Pays: Germany
ID NLM: 8703571
Informations de publication
Date de publication:
May 2022
May 2022
Historique:
received:
15
11
2021
accepted:
22
02
2022
pubmed:
2
3
2022
medline:
13
4
2022
entrez:
1
3
2022
Statut:
ppublish
Résumé
Toxoplasmosis is a zoonotic disease caused by the protozoan parasite Toxoplasma gondii. Infection in humans has usually been related to the consumption of raw, undercooked or cured meat. The aim of this study was to develop a droplet digital polymerase chain reaction (ddPCR)-based assay for the detection and quantification of T. gondii in meat samples. To optimize the ddPCR, T.gondii reference DNA aliquots at five known concentrations: 8000 cg/µl, 800 cg/µl, 80 cg/µl, 8 cg/µl were used. Moreover, results obtained by ddPCR and quantitative PCR (qPCR) were compared using 80 known samples (40 positive and 40 negative), as well as 171 unknown diaphragm tissue samples collected at slaughterhouses. The ddPCR showed a sensitivity of 97.5% and a specificity of 100%, with a detection limit of 8 genomic copy/µl of T. gondii. A nearly perfect agreement (κ = 0.85) was found between results obtained by ddPCR and qPCR for both positive and negative known samples analysed. On the 171 diaphragm tissue samples from field, 7.6% resulted positive by ddPCR and only 1.2% by qPCR. Therefore, this innovative method could be very useful for the detection of T. gondii in meat samples, aiming to prevent human infections.
Identifiants
pubmed: 35230549
doi: 10.1007/s00436-022-07477-9
pii: 10.1007/s00436-022-07477-9
pmc: PMC8993784
doi:
Substances chimiques
DNA, Protozoan
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
1467-1473Subventions
Organisme : Regione Campania
ID : This study was supported by the Regional Project "Control
Organisme : Regione Campania
ID : reduction of toxoplasmosis in animals
Organisme : Regione Campania
ID : humans- ToxoCamp"
Organisme : Regione Campania
ID : Campania Region
Organisme : Regione Campania
ID : Italy
Informations de copyright
© 2022. The Author(s).
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