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

Author:

Clark David J1ORCID,Moore Catherine M1,Flanagan Marc2,Bocxlaer Katrien Van3,Piperaki Evangelia-Theophano3,Yardley Vanessa3,Croft Simon L3,Tyson John2,Whitehouse Sam P2,O’Halloran Jonathan2,Krishna Sanjeev1,Staines Henry M1

Affiliation:

1. Centre for Diagnostics & Antimicrobial Resistance, Institute for Infection & Immunity, St George's University of London, Cranmer Terrace, London, SW17 0RE, UK

2. QuantuMDx, Newcastle upon Tyne, NE1 2JQ, UK

3. Infection & Immunity Department, Faculty of Infectious & Tropical Diseases, London School of Hygiene & Tropical Medicine, London, WC1E 7HT, UK

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.

Publisher

Future Science Ltd

Subject

General Biochemistry, Genetics and Molecular Biology,Biotechnology

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