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Métodos Terapêuticos e Terapias MTCI
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J Antimicrob Chemother ; 74(4): 854-864, 2019 04 01.
Artigo em Inglês | MEDLINE | ID: mdl-30649322

RESUMO

OBJECTIVES: Antimicrobial resistance genes (ARGs) can be transferred by means of mobile genetic elements, which play a critical role in the dissemination of resistance in the bacterial community. ARG transmission within mobile genetic elements has been reported in plasmids and transposons but less frequently in bacteriophages. Here, the bacteriophage fraction of seven human faecal samples was purified and deep-sequenced to detect the presence of ARGs in the phage particles. METHODS: Seven faecal samples (five from healthy individuals and two from a patient before and after receiving ciprofloxacin treatment) were used to extract phage DNA, which was purified and then sequenced in a MiSeq (Illumina). Generated reads were checked for quality and assembled, and then the generated contigs analysed with Kraken, PHASTER, VirSorter and Prokka. Some genes were also validated by quantitative PCR. RESULTS: Analysis of the purified phage DNA by Kraken identified from 4 to 266 viruses in the samples. The viral fraction corresponded mainly to the order Caudovirales, including phages from the Siphoviridae and Myoviridae families. Bacterial genes associated with antimicrobial resistance were detected in the viral DNA, as confirmed by quantitative PCR. Higher densities of ARG-carrying phage particles were observed in the post- versus pre-ciprofloxacin treatment sample. CONCLUSIONS: The finding of ARGs in phage particles supports the description of phages as mobile elements contributing to the dissemination of bacterial antibiotic resistance and suggests ciprofloxacin treatment may play a role in the release of ARG-carrying particles, thereby increasing resistance.


Assuntos
Antibacterianos/administração & dosagem , Bacteriófagos/isolamento & purificação , Ciprofloxacina/administração & dosagem , Farmacorresistência Bacteriana , Fezes/virologia , Genes Bacterianos , Voluntários Saudáveis , Adulto , Idoso , Bacteriófagos/classificação , Bacteriófagos/genética , Biota/efeitos dos fármacos , DNA Viral/química , DNA Viral/genética , DNA Viral/isolamento & purificação , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Pessoa de Meia-Idade , Myoviridae/classificação , Myoviridae/genética , Myoviridae/isolamento & purificação , Reação em Cadeia da Polimerase em Tempo Real , Siphoviridae/classificação , Siphoviridae/genética , Siphoviridae/isolamento & purificação
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