The Library
A nanoparticle-based method for culture-free bacterial DNA enrichment from whole blood
Tools
Hassan, Marwa, Ranzoni, Andrea and Cooper, Matthew A. (2018) A nanoparticle-based method for culture-free bacterial DNA enrichment from whole blood. Biosensors and Bioelectronics, 99 . pp. 150-155. doi:10.1016/j.bios.2017.07.057 ISSN 0956-5663.
Research output not available from this repository.
Request-a-Copy directly from author or use local Library Get it For Me service.
Official URL: http://dx.doi.org/10.1016/j.bios.2017.07.057
Abstract
Point-of-care (POC) diagnostics are one of the quick and sensitive detection approaches used in current clinical applications, but always face a performance tradeoff between time-to-result and assay sensitivity. One critical setting where these limitations are evident is the detection of sepsis, where 6–10 mL of whole blood may contain as little as one bacterial colony forming unit (cfu). The large sample volume, complex nature of the sample and low analyte concentration necessitates signal enhancement using culture-based or molecular amplification techniques. In the time-critical diagnosis of sepsis, waiting for up to 24 h to produce sufficient DNA for analysis is not possible. As a consequence, there is a need for integrated sample preparation methods that could enable shorter detection times, whilst maintaining high analytical performance. We report the development of a culture-free bacterial enrichment method to concentrate bacteria from whole blood in less than 3 h. The method relies on triple-enrichment steps to magnetically concentrate bacterial cells and their DNA with a 500-fold reduction in sample volume (from 10 to 0.02 mL). Using this sample preparation method, sensitive qPCR detection of the extracted S. aureus bacterial DNA was achieved with a detection limit of 5±0.58 cfu/mL within a total elapsed time of 4 h; much faster than conventional culture-based approaches. The method could be fully automated for integration into clinical practice for point-of-care or molecular detection of bacterial DNA from whole blood.
Item Type: | Journal Article | ||||||||
---|---|---|---|---|---|---|---|---|---|
Divisions: | Faculty of Science, Engineering and Medicine > Science > Life Sciences (2010- ) | ||||||||
Journal or Publication Title: | Biosensors and Bioelectronics | ||||||||
Publisher: | Elsevier BV | ||||||||
ISSN: | 0956-5663 | ||||||||
Official Date: | 15 January 2018 | ||||||||
Dates: |
|
||||||||
Volume: | 99 | ||||||||
Page Range: | pp. 150-155 | ||||||||
DOI: | 10.1016/j.bios.2017.07.057 | ||||||||
Status: | Peer Reviewed | ||||||||
Publication Status: | Published | ||||||||
Access rights to Published version: | Restricted or Subscription Access |
Request changes or add full text files to a record
Repository staff actions (login required)
View Item |