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Quantitative Real-Time PCR and Digital PCR to Evaluate Residual Quantity of HAV in Experimentally Depurated Mussels

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Abstract

Kinetics of hepatitis A virus (HAV) accumulation and depuration from mussels (Mytilus galloprovincialis) was studied in an experimental depuration system. Different parameters likely to influence the rate of virus accumulation and elimination were evaluated. Analyses were carried out by both real-time RT-qPCR and digital PCR. Results demonstrated that the animals start to concentrate the virus already after one hour and reach the maximum level of contamination in 6 h of experiment. With respect to depuration, HAV showed a rapid reduction of the concentration (89%) during the first 24–48 h of experiment and a very slow virus decrement in the following days with a 1% residual RNA at the ninth day of depuration. When process parameters likely to increase the depuration rate (presence of ozone, microalgal feeding, presence of lactic bacteria, pre-treatment with digestive enzymes) were tested, no significant differences in the kinetics were observed. Only treatment with pancreatin seemed to positively affect depuration in the first two days of the experiment.

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Funding

This work was supported by Grant RC015/IZSME from Italian Ministry of Health.

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Author notes

  1. MariaGrazia Amoroso and DeniseDi Concilio have equally contributed to the paper

Authors and Affiliations

  1. Unit of Virology, Department of Animal Health, Zooprofilactic and Experimental Institute of Southern Italy, Via Salute n. 2, Portici (Naples), Italy

    Maria Grazia Amoroso, Denise Di Concilio, Barbara Cioffi, Giorgio Galiero & Giovanna Fusco

  2. CAISIAL–Aquaculture division, University of Naples Federico II, Via Università n.133, Portici (Naples), Italy

    Antonio Luca Langellotti & Anna Martello

  3. Dipartimento Di Sicurezza Alimentare, Istituto Superiore Di Sanità, Nutrizione E Sanità Pubblica Veterinaria, Rep. Sicurezza Alimentare E Malattie Trasmesse Dagli Alimenti, Viale Regina Elena n. 299, Rome, Italy

    Elisabetta Suffredini, Loredana Cozzi & Simona Di Pasquale

  4. Department of Veterinary Medicine, University of Naples Federico II, Via Delpino n. 2, Naples, Italy

    Valeria Russo

  5. Department of Agriculture, University of Naples Federico II, Via Università n.133, Portici, Naples, Italy

    Gianluigi Mauriello

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  1. Maria Grazia Amoroso
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Correspondence to Maria Grazia Amoroso or Antonio Luca Langellotti.

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Amoroso, M.G., Di Concilio, D., Langellotti, A.L. et al. Quantitative Real-Time PCR and Digital PCR to Evaluate Residual Quantity of HAV in Experimentally Depurated Mussels. Food Environ Virol 13, 329–336 (2021). https://doi.org/10.1007/s12560-021-09470-4

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