TY - JOUR
T1 - Next-generation sequencing and maldi mass spectrometry in the study of multiresistant processed meat vancomycin-resistant enterococci (VRE)
AU - Sabença, Carolina
AU - de Sousa, Telma
AU - Oliveira, Soraia
AU - Viala, Didier
AU - Théron, Laetitia
AU - Chambon, Christophe
AU - Hébraud, Michel
AU - Beyrouthy, Racha
AU - Bonnet, Richard
AU - Caniça, Manuela
AU - Poeta, Patrícia
AU - Igrejas, Gilberto
N1 - Ação n.º: TC-14 / 2017
UID/QUI/50006/2020
UIDB/00211/2020
UMR1071
USC-2018
PY - 2020/5
Y1 - 2020/5
N2 - 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.
AB - 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.
KW - Antibiotic resistance
KW - Enterococcus spp
KW - MALDI-TOF MS
KW - Next-generation sequencing
KW - Processed meat
UR - http://www.scopus.com/inward/record.url?scp=85084118807&partnerID=8YFLogxK
U2 - 10.3390/biology9050089
DO - 10.3390/biology9050089
M3 - Article
C2 - 32349310
AN - SCOPUS:85084118807
VL - 9
JO - Biology
JF - Biology
IS - 5
M1 - 89
ER -