Preventing Chagas disease: A new RT-qPCR method for rapid and specific quantification of viable Trypanosoma cruzi for food safety.

Autor: de Oliveira AC; Centre for Valorisation of Amazonian Bioactive Compounds (CVACBA), Federal University of Pará, Av. Perimetral da Ciência km 01, 66.095-780 Belém, PA, Brazil., Thomaz-Soccol V; Department of Bioprocess Engineering and Biotechnology, Molecular Biology Laboratory, Federal University of Paraná, Curitiba, PR, Brazil., Tadra-Sfeir MZ; Department of Biochemistry, Centro Politécnico, Federal University of Paraná, Curitiba, PR, Brazil., de Souza EM; Department of Biochemistry, Centro Politécnico, Federal University of Paraná, Curitiba, PR, Brazil., Crespo-Lopez ME; Laboratory of Molecular Pharmacology, Institute of Biological Sciences, Federal University of Pará, Belém, PA, Brazil., Rogez H; Centre for Valorisation of Amazonian Bioactive Compounds (CVACBA), Federal University of Pará, Av. Perimetral da Ciência km 01, 66.095-780 Belém, PA, Brazil. Electronic address: herverogez@gmail.com.
Jazyk: angličtina
Zdroj: Food research international (Ottawa, Ont.) [Food Res Int] 2021 Jun; Vol. 144, pp. 110368. Date of Electronic Publication: 2021 Apr 17.
DOI: 10.1016/j.foodres.2021.110368
Abstrakt: Without standardized methods for rapidly detecting in food matrices viable T. cruzi, foodborne outbreaks remain neglected. In this work, a reverse-transcriptase real-time PCR (RT-qPCR) mRNA-based technique was developed for the rapid and specific detection and quantification of viable Trypanosoma cruzi in açai fruits and juice. The method uses specific primer targeting region on the cyt b gene. The maximum recovery rate of T. cruzi from inoculated açai juice was 82.50%. The limit of detection and quantification in açai juice was 10 parasites/mL for RT-qPCR (mRNA-based) and qPCR (DNA-based). The RT-qPCR efficiency was estimated at 97.27% with an R 2 of 0.994. The RT-qPCR was shown to be able to discriminate between viable and nonviable cells. This method provides a useful tool for rapid assessment of low concentrations of viable T. cruzi in naturally contaminated food samples, and can be applied industrially as a quality and security method.
(Copyright © 2021 Elsevier Ltd. All rights reserved.)
Databáze: MEDLINE
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