Rapid identification of intact bacterial resistance plasmids via optical mapping of single DNA molecules
Journal article, 2016

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.

Author

Lena Nyberg

Chalmers, Biology and Biological Engineering, Chemical Biology

Mahmood Saair Quaderi

Chalmers, Biology and Biological Engineering, Chemical Biology

Lund University

Gustav Emilsson

Chalmers, Physics, Bionanophotonics

N. Karami

Sahlgrenska Academy

E. Lagerstedt

Lund University

Vilhelm Müller

Chalmers, Biology and Biological Engineering, Chemical Biology

C. Noble

Lund University

S. Hammarberg

Lund University

A. Nilsson

Lund University

F. Sjoberg

Sahlgrenska Academy

Joachim Fritzsche

Chalmers, Physics, Chemical Physics

L. Sandegren

Uppsala University

T. Ambjornsson

Lund University

Fredrik Westerlund

Chalmers, Biology and Biological Engineering, Chemical Biology

Scientific Reports

2045-2322 (ISSN)

Vol. 6 30410

Driving Forces

Sustainable development

Innovation and entrepreneurship

Areas of Advance

Nanoscience and Nanotechnology

Life Science Engineering

DOI

10.1038/srep30410

PubMed

27460437

More information

Latest update

7/27/2018