Sponge Whirl-Pak Sampling Method and Droplet Digital RT-PCR Assay for Monitoring of SARS-CoV-2 on Surfaces in Public and Working Environments
Abstract
:1. Introduction
2. Materials and Methods
2.1. Sampling
2.2. Nucleic Acids Extraction
2.3. SARS-CoV-2 Detection by Real-Time RT-PCR
2.4. SARS-CoV-2 Detection in Environmental Samples by Droplet Digital RT-PCR (dd RT-PCR)
3. Results
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Sampling Location | District | Environmental Samples | Animal Swabs | Human Swabs |
---|---|---|---|---|
Buffaloes farm_1 | Caserta | 1 | 20 | 7 |
Buffaloes farm_2 | Caserta | 1 | 22 | 11 |
Buffaloes farm_3 | Caserta | 1 | 23 | 12 |
Buffaloes farm_4 | Caserta | 10 | 32 | 9 |
Cattle farm | Salerno | - | 10 | 1 |
Goat and cattle farm | Salerno | 2 | 17 | 2 |
Goat farm_1 | Salerno | 1 | 6 | - |
Goat farm_2 | Salerno | - | 10 | 1 |
Hen farm_1 | Salerno | 1 | 10 | 5 |
Hen farm_2 | Salerno | 1 | 11 | 10 |
Horse stall_1 | Salerno | 4 | 6 | 3 |
Horse stall_2 | Salerno | 6 | 14 | 4 |
Pig farm | Salerno | 1 | 9 | 7 |
Pig farm | Benevento | 1 | 16 | 6 |
Pig farm | Avellino | 2 | 21 | 2 |
Pigeon farm | Napoli | 1 | 12 | 2 |
Rabbits farm | Salerno | 1 | 7 | 11 |
Bus_1 | Salerno | 5 | - | - |
Bus_2 | Salerno | 5 | - | - |
Bus_3 | Salerno | 5 | - | - |
Bus_4 | Salerno | 5 | - | - |
Supermarket | Salerno | 14 | - | - |
Hotel receptive structure | Napoli | 9 | - | - |
Total | 77 | 246 | 93 |
Primers and Probes | Sequence | Concentrations |
---|---|---|
RdRp_SARSr-F | GTGARATGGTCATGTGTGGCGG | 600 nM |
RdRp_SARSr-R | CARATGTTAAASACACTATTAGCATA | 800 nM |
RdRP_SARSr-P1 | FAM-CCAGGTGGWACRTCATCMGGTGATGC-BBQ | 100 nM |
RdRp_SARSr-P2 | FAM-CAGGTGGAACCTCATCAGGAGATGC-BBQ | 100 nM |
E_Sarbeco_F | ACAGGTACGTTAATAGTTAATAGCGT | 400 nM |
E_Sarbeco_R | ATATTGCAGCAGTACGCACACA | 400 nM |
E_Sarbeco_P1 | FAM-ACACTAGCCATCCTTACTGCGCTTCG-BBQ | 200 nM |
N_Sarbeco_F | CACATTGGCACCCGCAATC | 600 nM |
N_Sarbeco_R | GAGGAACGAGAAGAGGCTTG | 800 nM |
N_Sarbeco_P | FAM-ACTTCCTCAAGGAACAACATTGCCA-BBQ | 200 nM |
Sampling Location | Municipality | N° of Samples | dd RT-PCR | ||
---|---|---|---|---|---|
RdRp | E Gene | N Gene | |||
Buffaloes farm_1 | Caserta | 1 | + | - | - |
Buffaloes farm_2 | Caserta | 1 | - | - | - |
Buffaloes farm_3 | Caserta | 1 | - | - | - |
Buffaloes farm_4 | Caserta | 1 | + | - | - |
Buffaloes farm_4 | Caserta | 2 | + | + | - |
Buffaloes farm_4 | Caserta | 7 | - | - | - |
Cattle farm | Salerno | 1 | - | - | - |
Goat and cattle farm | Salerno | 1 | - | - | - |
Goat farm_1 | Salerno | 1 | - | - | - |
Goat farm_2 | Salerno | 1 | + | - | - |
Hen farm_1 | Salerno | 1 | + | - | - |
Hen farm_2 | Salerno | 1 | - | - | - |
Horse stall_1 | Salerno | 2 | - | - | - |
Horse stall_1 | Salerno | 2 | + | + | - |
Horse stall_1 | Salerno | 2 | + | - | - |
Horse stall_2 | Salerno | 4 | - | - | - |
Pig farm | Salerno | 1 | - | - | - |
Pig farm | Benevento | 1 | - | - | - |
Pig farm | Avellino | 1 | - | - | - |
Pigeon farm | Napoli | 1 | - | - | - |
Rabbits farm | Salerno | 1 | - | + | - |
subtotal | 34 | 10 (29%) | 5 (15%) | 0 (0%) | |
Bus_1 | Salerno | 3 | - | - | - |
Bus_1 | Salerno | 1 | + | - | - |
Bus_1 | Salerno | 1 | + | + | - |
Bus_2 | Salerno | 1 | + | - | - |
Bus_2 | Salerno | 4 | - | - | - |
Bus_3 | Salerno | 1 | + | + | - |
Bus_3 | Salerno | 3 | - | - | - |
Bus_3 | Salerno | 1 | + | - | - |
Bus_4 | Salerno | 4 | - | - | - |
Bus_4 | Salerno | 1 | + | - | - |
subtotal | 20 | 6 (30%) | 2 (10%) | 0 (0%) | |
Supermarket | Salerno | 11 | - | - | - |
Supermarket | Salerno | 1 | - | + | - |
Supermarket | Salerno | 1 | - | - | + |
Supermarket | Salerno | 1 | + | - | - |
subtotal | 14 | 1 (7%) | 1 (7%) | 1 (7%) | |
Hotel receptive structure | Napoli | 9 | - | - | - |
subtotal | 9 | 0 (0%) | 0 (0%) | 0 (0%) | |
Total | 77 | 17 (22%) | 8 (10%) | 1 (1%) |
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Cardinale, D.; Tafuro, M.; Mancusi, A.; Girardi, S.; Capuano, F.; Proroga, Y.T.R.; Corrado, F.; D’Auria, J.L.; Coppola, A.; Rofrano, G.; et al. Sponge Whirl-Pak Sampling Method and Droplet Digital RT-PCR Assay for Monitoring of SARS-CoV-2 on Surfaces in Public and Working Environments. Int. J. Environ. Res. Public Health 2022, 19, 5861. https://doi.org/10.3390/ijerph19105861
Cardinale D, Tafuro M, Mancusi A, Girardi S, Capuano F, Proroga YTR, Corrado F, D’Auria JL, Coppola A, Rofrano G, et al. Sponge Whirl-Pak Sampling Method and Droplet Digital RT-PCR Assay for Monitoring of SARS-CoV-2 on Surfaces in Public and Working Environments. International Journal of Environmental Research and Public Health. 2022; 19(10):5861. https://doi.org/10.3390/ijerph19105861
Chicago/Turabian StyleCardinale, Davide, Maria Tafuro, Andrea Mancusi, Santa Girardi, Federico Capuano, Yolande Thérèse Rose Proroga, Federica Corrado, Jacopo Luigi D’Auria, Annachiara Coppola, Giuseppe Rofrano, and et al. 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" International Journal of Environmental Research and Public Health 19, no. 10: 5861. https://doi.org/10.3390/ijerph19105861