A comparison of DNA extraction methods used in the direct molecular detection of Burkholderia pseudomallei from blood.

Abstract

Melioidosis is caused by Burkholderia pseudomallei. Direct molecular detection from unamplified blood remains insensitive. Three different extraction methods-QIAamp UCP Pathogen Mini Kit, High Pure PCR template and MagNA Pure Pathogen Universal-were trialled using spiked human ethylenediaminetetraacetic acid blood. A type III secretion system 1 (TTSS-1) polymerase chain reaction was used for detection. The QIAamp UCP Pathogen Mini Kit performed best, with a limit of detection of 1.5×102cfu/ml. It is planned to use the QIAamp UCP Pathogen Mini Kit to do a larger study on blood collected from patients with melioidosis.