SORA

Advancing, promoting and sharing knowledge of health through excellence in teaching, clinical practice and research into the prevention and treatment of illness

An efficient and novel technology for the extraction of parasite genomic DNA from whole blood or culture

Clark, DJ; Moore, CM; Flanagan, M; Bocxlaer, KV; Piperaki, E-T; Yardley, V; Croft, SL; Tyson, J; Whitehouse, SP; O'Halloran, J; et al. Clark, DJ; Moore, CM; Flanagan, M; Bocxlaer, KV; Piperaki, E-T; Yardley, V; Croft, SL; Tyson, J; Whitehouse, SP; O'Halloran, J; Krishna, S; Staines, HM (2020) An efficient and novel technology for the extraction of parasite genomic DNA from whole blood or culture. Biotechniques, 68 (2). pp. 79-84. ISSN 1940-9818 https://doi.org/10.2144/btn-2019-0086
SGUL Authors: Staines, Henry Michael Moore, Catherine Margaret Krishna, Sanjeev Clark, David John

[img]
Preview
PDF Published Version
Available under License Creative Commons Attribution Non-commercial No Derivatives.

Download (613kB) | Preview
[img] Microsoft Word (.docx) (Title page) Published Version
Available under License Creative Commons Attribution Non-commercial No Derivatives.

Download (26kB)
[img]
Preview
PDF Accepted Version
Available under License Creative Commons Attribution Non-commercial No Derivatives.

Download (4MB) | Preview

Abstract

The aim of this study was to assess pathogen DNA extraction with a new spin column-based method (DNA-XT). DNA from either whole-blood samples spiked with Plasmodium falciparum or Leishmania donovani amastigote culture was extracted with DNA-XT and compared with that produced by a commercial extraction kit (DNeasy®). Eluates from large and small sample volumes were assessed by PCR and spectroscopy. Using a small volume (5 μl) of blood, the DNA-XT and DNeasy methods produced eluates with similar DNA concentrations (0.63 vs 1.06 ng/μl, respectively). The DNA-XT method produced DNA with lower PCR inhibition than DNeasy. The new technique was also twice as fast and required fewer plastics and manipulations but had reduced total recovered DNA compared with DNeasy.

Item Type: Article
Additional Information: © 2019 Dr Henry Staines This work is licensed under the Attribution-NonCommercial-NoDerivatives 4.0 Unported License. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-nd/4.0/
Keywords: DNA extraction, Leishmania, PCR, Plasmodium, blood, diagnostics, malaria, 06 Biological Sciences, 10 Technology, Bioinformatics
SGUL Research Institute / Research Centre: Academic Structure > Infection and Immunity Research Institute (INII)
Journal or Publication Title: Biotechniques
ISSN: 1940-9818
Language: eng
Dates:
DateEvent
February 2020Published
27 November 2019Published Online
30 October 2019Accepted
Publisher License: Creative Commons: Attribution-Noncommercial-No Derivative Works 4.0
Projects:
Project IDFunderFunder ID
304948Seventh Framework Programmehttp://dx.doi.org/10.13039/501100004963
MC_PC_14111 (2014 – 2015)Medical Research Councilhttp://dx.doi.org/10.13039/501100000265
204809/Z/16/ZWellcome Trusthttp://dx.doi.org/10.13039/100004440
PubMed ID: 31774301
Go to PubMed abstract
URI: http://openaccess.sgul.ac.uk/id/eprint/111368
Publisher's version: https://doi.org/10.2144/btn-2019-0086

Actions (login required)

Edit Item Edit Item