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

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((R))). Eluates from large and small sample volumes were assessed by PCR and spectroscopy. Using a small volume (5 mu l) of blood, the DNA-XT and DNeasy methods produced eluates with similar DNA concentrations (0.63 vs 1.06 ng/mu l, 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. METHOD SUMMARY DNA-XT, which is designed for small sample volumes, uses a 5-min detergent and enzymatic lysis step to release DNA from cells. Contaminating proteins and lipids are then bound to a matrix within a spin column during a 1-min centrifugation step while DNA passes directly through.