Rapid identification of intact bacterial resistance plasmids via optical mapping of single DNA molecules.
Sci Rep
; 6: 30410, 2016 07 27.
Article
em En
| MEDLINE
| ID: mdl-27460437
ABSTRACT
The rapid spread of antibiotic resistance - currently one of the greatest threats to human health according to WHO - is to a large extent enabled by plasmid-mediated horizontal transfer of resistance genes. Rapid identification and characterization of plasmids is thus important both for individual clinical outcomes and for epidemiological monitoring of antibiotic resistance. Toward this aim, we have developed an optical DNA mapping procedure where individual intact plasmids are elongated within nanofluidic channels and visualized through fluorescence microscopy, yielding barcodes that reflect the underlying sequence. The assay rapidly identifies plasmids through statistical comparisons with barcodes based on publicly available sequence repositories and also enables detection of structural variations. Since the assay yields holistic sequence information for individual intact plasmids, it is an ideal complement to next generation sequencing efforts which involve reassembly of sequence reads from fragmented DNA molecules. The assay should be applicable in microbiology labs around the world in applications ranging from fundamental plasmid biology to clinical epidemiology and diagnostics.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Plasmídeos
/
DNA
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Farmacorresistência Bacteriana
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Microfluídica
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Código de Barras de DNA Taxonômico
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Imagem Óptica
Tipo de estudo:
Diagnostic_studies
Idioma:
En
Ano de publicação:
2016
Tipo de documento:
Article