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Sponge Whirl-Pak Sampling Method and Droplet Digital RT-PCR Assay for Monitoring of SARS-CoV-2 on Surfaces in Public and Working Environments

Author

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  • Davide Cardinale

    (Centro di Referenza Nazionale per l’Analisi e Studio di Correlazione tra Ambiente, Animale e Uomo, Istituto Zooprofilattico Sperimentale del Mezzogiorno, Via Salute n. 2, 80055 Portici, Italy
    These authors contributed equally to this work.)

  • Maria Tafuro

    (Centro di Referenza Nazionale per l’Analisi e Studio di Correlazione tra Ambiente, Animale e Uomo, Istituto Zooprofilattico Sperimentale del Mezzogiorno, Via Salute n. 2, 80055 Portici, Italy
    These authors contributed equally to this work.)

  • Andrea Mancusi

    (Department of Food Security Coordination, Istituto Zooprofilattico Sperimentale del Mezzogiorno, Via Salute n. 2, 80055 Portici, Italy)

  • Santa Girardi

    (Department of Food Security Coordination, Istituto Zooprofilattico Sperimentale del Mezzogiorno, Via Salute n. 2, 80055 Portici, Italy)

  • Federico Capuano

    (Department of Food Security Coordination, Istituto Zooprofilattico Sperimentale del Mezzogiorno, Via Salute n. 2, 80055 Portici, Italy)

  • Yolande Thérèse Rose Proroga

    (Department of Food Security Coordination, Istituto Zooprofilattico Sperimentale del Mezzogiorno, Via Salute n. 2, 80055 Portici, Italy)

  • Federica Corrado

    (Centro di Referenza Nazionale per l’Analisi e Studio di Correlazione tra Ambiente, Animale e Uomo, Istituto Zooprofilattico Sperimentale del Mezzogiorno, Via Salute n. 2, 80055 Portici, Italy)

  • Jacopo Luigi D’Auria

    (Centro di Referenza Nazionale per l’Analisi e Studio di Correlazione tra Ambiente, Animale e Uomo, Istituto Zooprofilattico Sperimentale del Mezzogiorno, Via Salute n. 2, 80055 Portici, Italy)

  • Annachiara Coppola

    (Centro di Referenza Nazionale per l’Analisi e Studio di Correlazione tra Ambiente, Animale e Uomo, Istituto Zooprofilattico Sperimentale del Mezzogiorno, Via Salute n. 2, 80055 Portici, Italy)

  • Giuseppe Rofrano

    (Centro di Referenza Nazionale per l’Analisi e Studio di Correlazione tra Ambiente, Animale e Uomo, Istituto Zooprofilattico Sperimentale del Mezzogiorno, Via Salute n. 2, 80055 Portici, Italy)

  • Palmiero Volzone

    (Centro di Referenza Nazionale per l’Analisi e Studio di Correlazione tra Ambiente, Animale e Uomo, Istituto Zooprofilattico Sperimentale del Mezzogiorno, Via Salute n. 2, 80055 Portici, Italy)

  • Pio Galdi

    (Centro di Referenza Nazionale per l’Analisi e Studio di Correlazione tra Ambiente, Animale e Uomo, Istituto Zooprofilattico Sperimentale del Mezzogiorno, Via Salute n. 2, 80055 Portici, Italy)

  • Sabato De Vita

    (Centro di Referenza Nazionale per l’Analisi e Studio di Correlazione tra Ambiente, Animale e Uomo, Istituto Zooprofilattico Sperimentale del Mezzogiorno, Via Salute n. 2, 80055 Portici, Italy)

  • Alfonso Gallo

    (Centro di Referenza Nazionale per l’Analisi e Studio di Correlazione tra Ambiente, Animale e Uomo, Istituto Zooprofilattico Sperimentale del Mezzogiorno, Via Salute n. 2, 80055 Portici, Italy)

  • Elisabetta Suffredini

    (Department of Food Safety, Nutrition and Veterinary Public Health, Istituto Superiore di Sanità, Viale Regina Elena, 299, 00161 Rome, Italy)

  • Biancamaria Pierri

    (Centro di Referenza Nazionale per l’Analisi e Studio di Correlazione tra Ambiente, Animale e Uomo, Istituto Zooprofilattico Sperimentale del Mezzogiorno, Via Salute n. 2, 80055 Portici, Italy)

  • Pellegrino Cerino

    (Centro di Referenza Nazionale per l’Analisi e Studio di Correlazione tra Ambiente, Animale e Uomo, Istituto Zooprofilattico Sperimentale del Mezzogiorno, Via Salute n. 2, 80055 Portici, Italy)

  • Maria Morgante

    (Azienda Sanitaria Locale Avellino, 83100 Avellino, Italy)

Abstract

The SARS-CoV-2 can spread directly via saliva, respiratory aerosols and droplets, and indirectly by contact through contaminated objects and/or surfaces and by air. In the context of COVID-19 fomites can be an important vehicle of virus transmission and contribute to infection risk in public environments. The aim of the study was to analyze through surface sampling (sponge method) the presence of SARS-CoV-2 in public and working environments, in order to evaluate the risk for virus transmission. Seventy-seven environmental samples were taken using sterile sponges in 17 animal farms, 4 public transport buses, 1 supermarket and 1 hotel receptive structure. Furthermore, 246 and 93 swab samples were taken in the farms from animals and from workers, respectively. SARS-CoV-2 detection was conducted by real-time RT-PCR and by digital droplet RT-PCR (dd RT-PCR) using RdRp, gene E and gene N as targets. None of the human and animal swab samples were positive for SARS-CoV-2, while detection was achieved in 20 of the 77 sponge samples (26%) using dd RT-PCR. Traces of the RdRp gene, gene E and gene N were found in 17/77 samples (22%, average concentration 31.2 g.c./cm 2 , range 5.6 to 132 g.c./cm 2 ), 8/77 samples (10%, average concentration 15.1 g.c./cm 2 , range 6 to 36 g.c./cm 2 ), and in 1/77 (1%, concentration 7.2 g.c./cm 2 ). Higher detection rates were associated with sampling in animal farms and on public transport buses (32% and 30%) compared to the supermarket (21%) and the hotel (no detection). The result of the study suggests that the risk of contamination of surfaces with SARS-CoV-2 increases in environments in which sanitation strategies are not suitable and/or in highly frequented locations, such as public transportation. Considering the analytical methods, the dd RT-PCR was the only approach achieving detection of SARS-CoV-2 traces in environmental samples. Thus, dd RT-PCR emerges as a reliable tool for sensitive SARS-CoV-2 detection.

Suggested Citation

  • Davide Cardinale & Maria Tafuro & Andrea Mancusi & Santa Girardi & Federico Capuano & Yolande Thérèse Rose Proroga & Federica Corrado & Jacopo Luigi D’Auria & Annachiara Coppola & Giuseppe Rofrano & P, 2022. "Sponge Whirl-Pak Sampling Method and Droplet Digital RT-PCR Assay for Monitoring of SARS-CoV-2 on Surfaces in Public and Working Environments," IJERPH, MDPI, vol. 19(10), pages 1-14, May.
    • Handle: RePEc:gam:jijerp:v:19:y:2022:i:10:p:5861-:d:813444
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    References listed on IDEAS

    as
    1. Giuseppina Caggiano & Francesca Apollonio & Francesco Triggiano & Giusy Diella & Pasquale Stefanizzi & Marco Lopuzzo & Marilena D’Ambrosio & Nicola Bartolomeo & Giovanna Barbuti & Giovanni Trifone Sor, 2021. "SARS-CoV-2 and Public Transport in Italy," IJERPH, MDPI, vol. 18(21), pages 1-9, October.
    2. Simant Dube & Jian Qin & Ramesh Ramakrishnan, 2008. "Mathematical Analysis of Copy Number Variation in a DNA Sample Using Digital PCR on a Nanofluidic Device," PLOS ONE, Public Library of Science, vol. 3(8), pages 1-9, August.
    3. Perrine Marcenac & Geun Woo Park & Lindsey M. Duca & Nathaniel M. Lewis & Elizabeth A. Dietrich & Leslie Barclay & Azaibi Tamin & Jennifer L. Harcourt & Natalie J. Thornburg & Jared Rispens & Almea Ma, 2021. "Detection of SARS-CoV-2 on Surfaces in Households of Persons with COVID-19," IJERPH, MDPI, vol. 18(15), pages 1-10, August.
    4. Maria Teresa Montagna & Osvalda De Giglio & Carla Calia & Chrysovalentinos Pousis & Francesca Apollonio & Carmen Campanale & Giusy Diella & Marco Lopuzzo & Angelo Marzella & Francesco Triggiano & Vinc, 2021. "First Detection of Severe Acute Respiratory Syndrome Coronavirus 2 on the Surfaces of Tourist-Recreational Facilities in Italy," IJERPH, MDPI, vol. 18(6), pages 1-6, March.
    5. Giuseppina Caggiano & Francesco Triggiano & Francesca Apollonio & Giusy Diella & Marco Lopuzzo & Marilena D’Ambrosio & Fabrizio Fasano & Pasquale Stefanizzi & Giovanni Trifone Sorrenti & Pantaleo Maga, 2021. "SARS-CoV-2 RNA and Supermarket Surfaces: A Real or Presumed Threat?," IJERPH, MDPI, vol. 18(17), pages 1-9, September.
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