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.

Interactions between free-living amoebae and Cryptosporidium parvum: an experimental study

“…To evaluate the infectivity of oocysts, we adapted a qPCR method combined with HCT-8 cell culture (CC-qPCR) previously published by Kubina et al . [ 36 ]. Briefly, HCT-8 cells were cultured in 96-well plates (Thermo Fischer Scientific, Roskilde, Denmark) (2 × 10 4 cells per well) at +37 °C with 5% CO 2 to obtain about 90% confluence after 72 h of culture.…”