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Recovery Comparison of Two Virus Concentration Methods from Wastewater Using Cell Culture and Real-Time PCR

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Abstract

Enteric viruses are shed in the feces and may be present in environmental waters. Their detection in wastewater, even at low concentration, is a major challenge. In this study, recoveries of Echovirus 7 (EV7), virions and RNA in wastewater, using virus concentration methods were determined to evaluate the detection of infectious viruses and the possibility of recovering viral genomes. Two virus concentration methods, PEG precipitation method and two-phase separation method, were applied to recovery experiments of EV7–virions from wastewater, in parallel with recovery experiments of EV7 RNA. The titration of EV7 virions was carried out by cell culture using human rhabdomyosarcoma tumor tissue and the EV7 RNA quantification was performed by real-time PCR. The mean recovery yields of EV7 virions using the PEG precipitation method and the two-phase separation method were 78.5 ± 10.99 and 83.1 ± 0.28 %, respectively. Besides, EV7 RNA recoveries obtained using the PEG precipitation method were four times higher than those using the two-phase separation method. According to our results, the two methods enable to concentrate both infectious viruses and viral genomes. Moreover, considering the protocol time and cost together with the ratio of the EV7 virion recovery to the EV7 RNA recovery, the two-phase separation method (83.1/2.71 %, or 30.6) seems to be more appropriate for selective concentration of viral virions than the PEG precipitation method (78.5/10.33 %, or 7.6).

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Acknowledgments

The authors thank Prof Albert Bosch from the University of Barcelona (Barcelona, Spain) for providing us with the internal control.

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Authors and Affiliations

  1. Medical Virology & BSL3 Laboratory, Institut Pasteur du Maroc, 1, Place Louis Pasteur, 20360, Casablanca, Morocco

    Hasna Amdiouni, Abdellah Faouzi & Jalal Nourlil

  2. Laboratory of Physiology and Molecular Genetics, Faculty of Sciences, Aîn Chock, University Hassan II, Km 8 Road of Eljadida BP, 5366, Casablanca, Morocco

    Hasna Amdiouni & Abdelaziz Soukri

  3. Department of Food Hygiene and Environmental Health, Faculty of Veterinary Medicine, University of Helsinki, 00014, Helsinki, Finland

    Leena Maunula

  4. Laboratory of Microbiology and Food Safety, 1, Place Louis Pasteur, 20360, Casablanca, Morocco

    Kawtar Hajjami

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  1. Hasna Amdiouni
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  2. Leena Maunula
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  3. Kawtar Hajjami
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  4. Abdellah Faouzi
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  5. Abdelaziz Soukri
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  6. Jalal Nourlil
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Correspondence to Hasna Amdiouni.

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Amdiouni, H., Maunula, L., Hajjami, K. et al. Recovery Comparison of Two Virus Concentration Methods from Wastewater Using Cell Culture and Real-Time PCR. Curr Microbiol 65, 432–437 (2012). https://doi.org/10.1007/s00284-012-0174-8

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  • DOI: https://doi.org/10.1007/s00284-012-0174-8

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