Detection of Infectious Cryptosporidium parvum Oocysts from Lamb’s Lettuce: CC-qPCR’s Intake

L’émergence de nouveaux modes alimentaires, la quête du manger « sain et naturel », incitent les consommateurs à manger, notamment des aliments frais comme les fruits et légumes. Parallèlement, un nombre croissant de maladies d’origine alimentaire causées par un large éventail d’agents pathogènes (dont les parasites protozoaires tels que Giardia duodenalisCryptosporidium spp. et Cyclospora cayetanensis) liés à la consommation d’aliment crus ou peu cuits frais a été signalé. Selon les rapports de l’EFSA-ECDC, Cryptosporidium spp. est considéré comme l’un des agents étiologiques à prendre en compte dans le domaine alimentaire. Ainsi, l’évaluation de l’exposition du consommateur au danger Cryptosporidium est cruciale pour la santé publique. Nous avons développé une méthode de détection de Cryptosporidium capable d’évaluer le risque d’infection à Cryptosporidium par la consommation d’aliments contaminés .

Nombre d’oocystes inoculés
par 30 g de mâche
10,000
1000
100
10
1
Nombre de lot
d’oocystes utilisé
2
2
2
2
2
Réplicat en culture cellulaire
6
6
6
6
6
Réplicat de qPCR
2
2
2
2
2
Nombre de puits détectés positifs
avec cellules
18/18
12/12
18/18
8/12
3/12
sans cellules
18/18
12/12
9/18
0/12
0/12
Cq obtenu avec cellules
25.79
29.16
31.91
34.18
35.40
ET obtenu avec cellules
2.38
1.74
2.52
1.58
0.87
Cq obtenu sans cellules
30.82
33.94
34.86
N/D
N/D
ET obtenu sans cellules
1.40
1.26
1.77
N/D
N/D

Detection of Infectious Cryptosporidium parvum Oocysts from Lamb’s Lettuce: CC-qPCR’s Intake

Abstract
Cryptosporidium spp. is responsible for several food and waterborne disease outbreaks worldwide. Healthier lifestyles attract consumers to eat, notably, fresh food like fruits and vegetables. The consumption of raw or under-cooked food increases the risk of foodborne transmission of Cryptosporidiosis. The assessment of the consumer’s exposure to Cryptosporidium danger is crucial for public health. Still, the standardized method to detect this parasite in fresh leafy greens and berry fruits has only been available since 2016 and suffers from weaknesses. Consequently, in this study, we propose a method with minimum processing steps that combines cell culture and the quantitative PCR (CC-qPCR) for detecting infectious C. parvum oocysts recovered from lamb’s lettuce. This CC-qPCR is a rapid and easy method that can detect up to one oocyst, whereas it is undetectable by classic qPCR.

Keywords: Cryptosporidium; detection; fresh vegetables; in vitro method; oocysts infectivity.

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