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Real-time PCR versus digital PCR in diagnostics of food-borne parasites

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RESLOVÁ Nikol KAŠNÝ Martin SLANÝ Michal MONTEIRO Silvia SANTOS Ricardo

Rok publikování 2016
Druh Konferenční abstrakty
Fakulta / Pracoviště MU

Přírodovědecká fakulta

Citace
Popis In recent years, an increased number of reported cases of human infections by parasitic agents present in foodstuffs is recorded in Europe. Among the dominant pathogens are documented protozoa - Giardia intestinalis and Toxoplasma gondii. Cysts of G. intestinalis can survive long term in a cool and moist environment and contaminate e.g. fruit and vegetables growing in soil. Whereas T. gondii is an obligate intracellular parasite of warm-blooded animals, which forms tissue cysts in nervous and muscle tissue of the intermediate host; during the meat processing might be contaminated also tissue of uninfected hosts. Moreover, meat inspection of T. gondii at the slaughterhouses is not legislatively established. The detection possibilities of G. intestinalis and T. gondii were tested using two molecular methods - quantitative real-time PCR (qPCR) and digital PCR (dPCR). While qPCR is a commonly used method for the detection of food-borne parasites, dPCR represents a new potentially fast, accurate and sensitive platform on this field. dPCR DNA chips enable to screen the absolute number of specifically amplified DNA molecules, which means that the high detection limit of pathogens could be reach. The aim of our study is to compare the efficiency of two methodological approaches adopted for detection of parasites in artificially contaminated samples of meat and vegetables by defined amounts of G. intestinalis (trofozoits) and T. gondii (oocysts).
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