Detecting Carbapenemases in Animal and Food Samples by Droplet Digital PCR
Maria Carelli,
Francesca Griggio,
Marina Mingoia,
Cristiana Garofalo,
Vesna Milanović,
Nicola Pozzato,
Francesca Leoni,
Laura Veschetti,
Giovanni Malerba,
Angela Sandri,
Cristina Patuzzo,
Serena Simoni,
Maria M. Lleo,
Carla Vignaroli
Affiliations
Maria Carelli
Department of Diagnostics and Public Health, University of Verona, 37134 Verona, Italy
Francesca Griggio
Centro Piattaforme Tecnologiche, University of Verona, 37134 Verona, Italy
Marina Mingoia
Department of Biomedical Sciences and Public Health, Polytechnic University of Marche, 60121 Ancona, Italy
Cristiana Garofalo
Department of Agricultural, Food and Environmental Sciences, Polytechnic University of Marche, 60121 Ancona, Italy
Vesna Milanović
Department of Agricultural, Food and Environmental Sciences, Polytechnic University of Marche, 60121 Ancona, Italy
Nicola Pozzato
Laboratorio di Diagnostica Clinica e Sierologia di Piano, Istituto Zooprofilattico Sperimentale delle Venezie, 37060 Buttapietra, Italy
Francesca Leoni
Laboratorio Nazionale di Riferimento (LNR) per il Controllo delle Contaminazioni Batteriche dei Molluschi Bivalvi, Istituto Zooprofilattico Sperimentale dell’Umbria e delle Marche “Togo Rosati”, 60121 Ancona, Italy
Laura Veschetti
Department of Neurosciences, Biomedicine and Movement Sciences, University of Verona, 37134 Verona, Italy
Giovanni Malerba
Department of Neurosciences, Biomedicine and Movement Sciences, University of Verona, 37134 Verona, Italy
Angela Sandri
Department of Diagnostics and Public Health, University of Verona, 37134 Verona, Italy
Cristina Patuzzo
Department of Neurosciences, Biomedicine and Movement Sciences, University of Verona, 37134 Verona, Italy
Serena Simoni
Department of Life and Environmental Sciences, Polytechnic University of Marche, 60121 Ancona, Italy
Maria M. Lleo
Department of Diagnostics and Public Health, University of Verona, 37134 Verona, Italy
Carla Vignaroli
Department of Life and Environmental Sciences, Polytechnic University of Marche, 60121 Ancona, Italy
Background: The presence of carbapenemase-producing bacteria (CPB) in animal hosts and along the food chain may result in the development of reservoirs for human infections. Several CPB strains isolated from animals have been reported, suggesting that transmission and dissemination of the corresponding genes between humans and animals may occur. Animal and food samples have complex backgrounds that hinder the detection of CPB present in low concentrations by standard detection procedures. Methods: We evaluated the possibility of detecting blaKPC, blaVIM, and blaOXA-48-like carbapenemases in 286 animal and food samples (faeces from farm and companion animals, raw meat, bivalve molluscs) by culture-based and standard molecular methods and by ddPCR. Results: The proposed ddPCR managed to detect the target genes, also in samples resulting negative to standard methods. While the presence of blaKPC and blaVIM was detected in few samples (~3%), one third of the samples (n = 94/283) carried different variants of blaOXA-48-like genes. Conclusion: A specific and sensitive method such as ddPCR could be suitable to evaluate the current veterinarian and environmental situation and to assess the dynamic transmission and persistence of CPB between animals and humans and vice versa.
