Next-Generation Sequencing and MALDI Mass Spectrometry in the Study of Multiresistant Processed Meat Vancomycin-Resistant Enterococci (VRE)

Biology

Racha Beyrouthy¹  Richard Bonnet¹  Patrícia Poeta²  Carolina Sabença³  Soraia Oliveira³  Gilberto Igrejas³  Telma de Sousa³  Michel Hébraud⁴  Didier Viala⁴  Christophe Chambon⁴  Laetitia Théron⁵  Manuela Caniça⁶ 

[1] Centre National de Référence de la Résistance aux Antibiotiques, Centre Hospitalier Universitaire, 63003 Clermont-Ferrand, France;Department of Animal and Veterinary Science, University of Trás-os-Montes and Alto Douro (UTAD), 5000-801 Vila Real, Portugal;Department of Genetics and Biotechnology, University of Trás-os-Montes and Alto Douro (UTAD), 5000-801 Vila Real, Portugal;INRAE, Plateforme d’Exploration du Métabolisme, composante protéomique (PFEMcp), 63122 Saint-Genès Champanelle, France;INRAE, UR0370 Qualité des Produits Animaux (QuaPA), 63122 Saint-Genès Champanelle, France;National Reference Laboratory of Antibiotic Resistances and Healthcare Associated Infections, National Institute of Health Dr. Ricardo Jorge, 1649-016 Lisbon, Portugal;

关键词: Enterococcus spp.;    processed meat;    antibiotic resistance;    next-generation sequencing;    MALDI-TOF MS;   

DOI  :  10.3390/biology9050089

来源: DOAJ

【 摘 要 】

Vancomycin-resistant enterococci (VRE), due to their intrinsic resistance to various commonly used antibiotics and their malleable genome, make the treatment of infections caused by these bacteria less effective. The aims of this work were to characterize isolates of Enterococcus spp. that originated from processed meat, through phenotypic and genotypic techniques, as well as to detect putative antibiotic resistance biomarkers. The 19 VRE identified had high resistance to teicoplanin (89%), tetracycline (94%), and erythromycin (84%) and a low resistance to kanamycin (11%), gentamicin (11%), and streptomycin (5%). Based on a Next-Generation Sequencing NGS technique, most isolates were vanA-positive. The most prevalent resistance genes detected were erm(B) and aac(6')-Ii, conferring resistance to the classes of macrolides and aminoglycosides, respectively. MALDI-TOF mass spectrometry (MS) analysis detected an exclusive peak of the Enterococcus genus at m/z (mass-to-charge-ratio) 4428 ± 3, and a peak at m/z 6048 ± 1 allowed us to distinguish Enterococcus faecium from the other species. Several statistically significant protein masses associated with resistance were detected, such as peaks at m/z 6358.27 and m/z 13237.3 in ciprofloxacin resistance isolates. These results reinforce the relevance of the combined and complementary NGS and MALDI-TOF MS techniques for bacterial characterization.

【 授权许可】

Unknown