A rapid and sensitive method to detect Toxoplasma gondii oocysts in soil samples.

Autor: Escotte-Binet S; Université de Reims Champagne Ardenne, ESCAPE EA 7510, CAP SANTE, 51097, Reims, France. Electronic address: sandie.escotte@univ-reims.fr., Da Silva AM; Université de Reims Champagne Ardenne, ESCAPE EA 7510, CAP SANTE, 51097, Reims, France. Electronic address: abdou_malik.da_silva@univ-fcomte.fr., Cancès B; Université de Reims Champagne Ardenne, GEGENAA EA 3795, CONDORCET, 51097 Reims, France. Electronic address: benjamin.cances@univ-reims.fr., Aubert D; Université de Reims Champagne Ardenne, ESCAPE EA 7510, CAP SANTE, 51097, Reims, France; CHU Reims, Hôpital Maison Blanche, Centre National de Référence de la Toxoplasmose, CRB Toxoplasma, Laboratoire de Parasitologie-Mycologie, 51097, Reims, France. Electronic address: daubert@chu-reims.fr., Dubey J; United States Department of Agriculture, Agricultural Research Service, Beltsville Agricultural Research Center, Animal Parasitic Diseases Laboratory, Building 1001, Beltsville, MD, 20705 2350, USA. Electronic address: dubey@ars.usda.gov., La Carbona S; ACTALIA, Food Safety Department, 310 Rue Popielujko, 50000, Saint-Lô, France. Electronic address: s.lacarbona@actalia.eu., Villena I; Université de Reims Champagne Ardenne, ESCAPE EA 7510, CAP SANTE, 51097, Reims, France; CHU Reims, Hôpital Maison Blanche, Centre National de Référence de la Toxoplasmose, CRB Toxoplasma, Laboratoire de Parasitologie-Mycologie, 51097, Reims, France. Electronic address: ivillena@chu-reims.fr., Poulle ML; Université de Reims Champagne Ardenne, ESCAPE EA 7510, CAP SANTE, 51097, Reims, France; Université de Reims Champagne Ardenne, CERFE, 08240, Boult-aux-Bois, France. Electronic address: marie-lazarine.poulle@univ-reims.fr.
Jazyk: angličtina
Zdroj: Veterinary parasitology [Vet Parasitol] 2019 Oct; Vol. 274, pp. 108904. Date of Electronic Publication: 2019 Aug 17.
DOI: 10.1016/j.vetpar.2019.07.012
Abstrakt: Documenting the extent of soil contamination by Toxoplasma gondii oocysts is a key issue to prevent the worldwide infection caused by this protozoan. Our aim was to improve the practicability and sensitivity of a low-cost method to detect T. gondii DNA in soil samples developed a few years ago. Various parameters of the reference protocol were modified to determine their effect on the detection of T. gondii DNA in soil samples ("natural soil" and "sand") spiked with oocysts. We tested i) filtration using stomacher bags, ii) Tween 80, Tween 20, SDS and Triton X100 as dispersion solutions, iii) sucrose solution, zinc chloride solution, Optiprep and Percoll as density gradients, iv) freeze/thaw versus mechanical grinding as lysis methods, and v) Qiagen versus Fastprep as extraction kits The optimized protocol is quicker and easier to use than the previous one, and includes the following items: 0.1% Tween80/PBS for dispersion, sucrose solution for flotation, mechanical grinding, and FastDNA spin kit for extraction. It accurately detects T. gondii DNA in both fresh and frozen soil samples and displays a detection limit below 1 oocyst/g of fresh soil.
(Copyright © 2019 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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