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1.
Artigo em Inglês | MEDLINE | ID: mdl-27993847

RESUMO

Sixteen different sequence types (STs) of Escherichia coli isolates from a commercial swine farm in China were confirmed to coharbor the carbapenem resistance gene blaNDM-5 and the colistin resistance gene mcr-1 Whole-genome sequencing revealed that blaNDM-5 and mcr-1 were located on a 46-kb IncX3 plasmid and a 32-kb IncX4 plasmid, respectively. The two plasmids can transfer together with a low fitness cost, which might explain the presence of various STs of E. coli coharboring blaNDM-5 and mcr-1.


Assuntos
Farmacorresistência Bacteriana Múltipla/genética , Infecções por Escherichia coli/veterinária , Proteínas de Escherichia coli/genética , Escherichia coli/genética , Doenças dos Suínos/epidemiologia , beta-Lactamases/genética , Animais , Antibacterianos/farmacologia , Carbapenêmicos/farmacologia , China/epidemiologia , Colistina/farmacologia , Escherichia coli/efeitos dos fármacos , Escherichia coli/crescimento & desenvolvimento , Escherichia coli/isolamento & purificação , Infecções por Escherichia coli/epidemiologia , Infecções por Escherichia coli/microbiologia , Proteínas de Escherichia coli/metabolismo , Expressão Gênica , Transferência Genética Horizontal , Aptidão Genética , Genótipo , Plasmídeos/química , Plasmídeos/genética , Plasmídeos/metabolismo , Suínos , Doenças dos Suínos/microbiologia , beta-Lactamases/metabolismo
2.
J Microbiol Methods ; 153: 24-30, 2018 10.
Artigo em Inglês | MEDLINE | ID: mdl-30099005

RESUMO

Salmonella enterica serovar Indiana (S. Indiana) was the most frequently reported foodborne pathogen, which has a broad host range including poultry, swine, and humans. Traditional methods used for the detection of S. Indiana from contaminated food products are time-consuming and labor-intensive. Therefore, rapid detection methods with high sensitivity and specificity are vitally important to prevent the spread of S. Indiana. In this study, we developed a nearly instrument-free, simple molecular method which incorporates cross-priming amplification (CPA) combined with a nucleic acid detection strip (NADS) for sensitive detection of S. Indiana. A set of CPA primers was designed based on S. Indiana specific nucleotide sequences and the specificity of CPA-NADS was tested against 42 bacterial strains. The results showed that this method was highly specific for detection of S. Indiana. The sensitivity of CPA-NADS was evaluated and compared with that of the serovar-specific PCR method and the real-time PCR method. The limit of detection of the CPA method was 8.997 fg/µL for genomic DNA and 6.2 × 101 CFU/mL for bacteria in pure culture. An application of the CPA assay was conducted with 90 inoculated specimens by S. Indiana. The accuracy of CPA-NADS was consistent with the results of the traditional culture-based methods in inoculated specimens. This method showed a higher sensitivity than the serovar-specific PCR method did and was more convenient to perform. In conclusion, we demonstrated that the CPA-NADS system offers high specificity, sensitivity, rapidity, and a simple detection tool for screening S. Indiana.


Assuntos
DNA Bacteriano/isolamento & purificação , Análise de Alimentos/métodos , Técnicas de Amplificação de Ácido Nucleico/métodos , Salmonella enterica/isolamento & purificação , Sorogrupo , Primers do DNA/genética , Contaminação de Alimentos/análise , Microbiologia de Alimentos , Limite de Detecção , Fitas Reagentes , Reação em Cadeia da Polimerase em Tempo Real , Reprodutibilidade dos Testes , Sensibilidade e Especificidade
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