Development of a qRT-PCR method to assess ...
Document type :
Article dans une revue scientifique
Title :
Development of a qRT-PCR method to assess the viability of Giardia intestinalis cysts, Cryptosporidium spp. and Toxoplasma gondii oocysts
Author(s) :
Travaillé, Emmanuelle [Auteur]
LABÉO, Pôle d’analyses et de recherche de Normandie [LABÉO]
La Carbona, Stéphanie [Auteur]
ACTALIA [Villers-Bocage]
Gargala, Gilles [Auteur]
Protozooses Transmises par l'Alimentation (Cryptosporidiose, Giardose et Toxoplasmose) : Mode de Contamination et Pathogénie (PROTAL) - EA 3800 [PROTAL]
Voutquenne-Nazabadioko, Laurence [Auteur]
Protozooses Transmises par l'Alimentation (Cryptosporidiose, Giardose et Toxoplasmose) : Mode de Contamination et Pathogénie (PROTAL) - EA 3800 [PROTAL]
Centre Hospitalier Universitaire de Reims [CHU Reims]
Guyot, Karine [Auteur]
Centre d’Infection et d’Immunité de Lille - INSERM U 1019 - UMR 9017 - UMR 8204 [CIIL]
Dumètre, Aurélien [Auteur]
Infections Parasitaires : Transmission, Physiopathologie et Thérapeutiques [IP-TPT]
Villena, Isabelle [Auteur]
Centre Hospitalier Universitaire de Reims [CHU Reims]
Protozooses Transmises par l'Alimentation (Cryptosporidiose, Giardose et Toxoplasmose) : Mode de Contamination et Pathogénie (PROTAL) - EA 3800 [PROTAL]
Houssin, Maryline [Auteur correspondant]
LABÉO, Pôle d’analyses et de recherche de Normandie [LABÉO]
LABÉO, Pôle d’analyses et de recherche de Normandie [LABÉO]
La Carbona, Stéphanie [Auteur]
ACTALIA [Villers-Bocage]
Gargala, Gilles [Auteur]
Protozooses Transmises par l'Alimentation (Cryptosporidiose, Giardose et Toxoplasmose) : Mode de Contamination et Pathogénie (PROTAL) - EA 3800 [PROTAL]
Voutquenne-Nazabadioko, Laurence [Auteur]
Protozooses Transmises par l'Alimentation (Cryptosporidiose, Giardose et Toxoplasmose) : Mode de Contamination et Pathogénie (PROTAL) - EA 3800 [PROTAL]
Centre Hospitalier Universitaire de Reims [CHU Reims]
Guyot, Karine [Auteur]
Centre d’Infection et d’Immunité de Lille - INSERM U 1019 - UMR 9017 - UMR 8204 [CIIL]
Dumètre, Aurélien [Auteur]
Infections Parasitaires : Transmission, Physiopathologie et Thérapeutiques [IP-TPT]
Villena, Isabelle [Auteur]
Centre Hospitalier Universitaire de Reims [CHU Reims]
Protozooses Transmises par l'Alimentation (Cryptosporidiose, Giardose et Toxoplasmose) : Mode de Contamination et Pathogénie (PROTAL) - EA 3800 [PROTAL]
Houssin, Maryline [Auteur correspondant]
LABÉO, Pôle d’analyses et de recherche de Normandie [LABÉO]
Journal title :
Food Control
Pages :
359-365
Publisher :
Elsevier
Publication date :
2016-01
ISSN :
0956-7135
English keyword(s) :
Giardia intestinalis
Cryptosporidium parvum
Toxoplasma gondii
Reverse transcription qPCR
Viability
Cryptosporidium parvum
Toxoplasma gondii
Reverse transcription qPCR
Viability
HAL domain(s) :
Sciences du Vivant [q-bio]/Ecologie, Environnement/Santé
Sciences du Vivant [q-bio]/Médecine humaine et pathologie/Maladies infectieuses
Sciences du Vivant [q-bio]/Microbiologie et Parasitologie
Sciences du Vivant [q-bio]/Médecine humaine et pathologie/Maladies infectieuses
Sciences du Vivant [q-bio]/Microbiologie et Parasitologie
English abstract : [en]
This study evaluates the viability of Giardia intestinalis cysts, Cryptosporidium spp. and Toxoplasma gondii oocysts, which may be found in water and food matrices subjected to contaminated water. Viability is a key factor ...
Show more >This study evaluates the viability of Giardia intestinalis cysts, Cryptosporidium spp. and Toxoplasma gondii oocysts, which may be found in water and food matrices subjected to contaminated water. Viability is a key factor to be considered in order to assess parasite infectivity. People living in developing countries are particularly at risk of contamination by these pathogens, which can cause severe diseases particularly in immunocompromised people. In this report, we describe methods combining the mechanical rupture of (oo)cysts and mRNA extraction in order to determine their viability by qRT-PCR. The targeted genes are beta-giardin, hsp70, and SporoSAG for the detection of G. intestinalis, Cryptosporidium spp. and T. gondii respectively. The proposed method is compared to in vivo infectivity tests specific of each parasite. Then, this qRT-PCR method is applied directly on parasite suspensions and subsequently on basil experimentally contaminated with the parasites. For each protozoan, the detection limit of the qRT-PCR method is 2 parasites per reaction (2 μL mRNA analyzed) on parasite suspensions, and 3 parasites per gram of basil matrix. Compared to in vivo methods, qRT-PCR can detect viable but not infectious parasites. This qRT-PCR method could represent an attractive alternative as a specific and sensitive tool for: i) rapid assessment of the risk of human contamination by G. intestinalis, Cryptosporidium spp. and T. gondii parasites; ii) evaluation of the efficiency of industrial process.Show less >
Show more >This study evaluates the viability of Giardia intestinalis cysts, Cryptosporidium spp. and Toxoplasma gondii oocysts, which may be found in water and food matrices subjected to contaminated water. Viability is a key factor to be considered in order to assess parasite infectivity. People living in developing countries are particularly at risk of contamination by these pathogens, which can cause severe diseases particularly in immunocompromised people. In this report, we describe methods combining the mechanical rupture of (oo)cysts and mRNA extraction in order to determine their viability by qRT-PCR. The targeted genes are beta-giardin, hsp70, and SporoSAG for the detection of G. intestinalis, Cryptosporidium spp. and T. gondii respectively. The proposed method is compared to in vivo infectivity tests specific of each parasite. Then, this qRT-PCR method is applied directly on parasite suspensions and subsequently on basil experimentally contaminated with the parasites. For each protozoan, the detection limit of the qRT-PCR method is 2 parasites per reaction (2 μL mRNA analyzed) on parasite suspensions, and 3 parasites per gram of basil matrix. Compared to in vivo methods, qRT-PCR can detect viable but not infectious parasites. This qRT-PCR method could represent an attractive alternative as a specific and sensitive tool for: i) rapid assessment of the risk of human contamination by G. intestinalis, Cryptosporidium spp. and T. gondii parasites; ii) evaluation of the efficiency of industrial process.Show less >
Language :
Anglais
Peer reviewed article :
Oui
Audience :
Internationale
Popular science :
Non
Source :