An efficient and novel technology for the extraction of parasite genomic DNA from whole blood or culture
David J Clark,
Catherine M Moore,
Marc Flanagan,
Katrien Van Bocxlaer,
Evangelia-Theophano Piperaki,
Vanessa Yardley,
Simon L Croft,
John Tyson,
Sam P Whitehouse,
Jonathan O’Halloran,
Sanjeev Krishna,
Henry M Staines
Affiliations
David J Clark
1Centre for Diagnostics & Antimicrobial Resistance, Institute for Infection & Immunity, St George's University of London, Cranmer Terrace, London, SW17 0RE, UK
Catherine M Moore
1Centre for Diagnostics & Antimicrobial Resistance, Institute for Infection & Immunity, St George's University of London, Cranmer Terrace, London, SW17 0RE, UK
Marc Flanagan
2QuantuMDx, Newcastle upon Tyne, NE1 2JQ, UK
Katrien Van Bocxlaer
3Infection & Immunity Department, Faculty of Infectious & Tropical Diseases, London School of Hygiene & Tropical Medicine, London, WC1E 7HT, UK
Evangelia-Theophano Piperaki
3Infection & Immunity Department, Faculty of Infectious & Tropical Diseases, London School of Hygiene & Tropical Medicine, London, WC1E 7HT, UK
Vanessa Yardley
3Infection & Immunity Department, Faculty of Infectious & Tropical Diseases, London School of Hygiene & Tropical Medicine, London, WC1E 7HT, UK
Simon L Croft
3Infection & Immunity Department, Faculty of Infectious & Tropical Diseases, London School of Hygiene & Tropical Medicine, London, WC1E 7HT, UK
John Tyson
2QuantuMDx, Newcastle upon Tyne, NE1 2JQ, UK
Sam P Whitehouse
2QuantuMDx, Newcastle upon Tyne, NE1 2JQ, UK
Jonathan O’Halloran
2QuantuMDx, Newcastle upon Tyne, NE1 2JQ, UK
Sanjeev Krishna
1Centre for Diagnostics & Antimicrobial Resistance, Institute for Infection & Immunity, St George's University of London, Cranmer Terrace, London, SW17 0RE, UK
Henry M Staines
1Centre for Diagnostics & Antimicrobial Resistance, Institute for Infection & Immunity, St George's University of London, Cranmer Terrace, London, SW17 0RE, UK
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.
