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

Georgia Mataira; Fleur Francis; Jaimie Frazer; Robert Norton

doi:10.1093/trstmh/trae020

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

Trans R Soc Trop Med Hyg. 2024 Apr 8:trae020. doi: 10.1093/trstmh/trae020. Online ahead of print.

Authors

Georgia Mataira¹, Fleur Francis¹, Jaimie Frazer¹, Robert Norton^{1

2}

Affiliations

¹ Pathology Queensland, Townsville University Hospital, Douglas, Townsville, Queensland 4814, Australia.
² Faculty of Medicine, University of Queensland, Brisbane, Australia.

PMID: 38586993
DOI: 10.1093/trstmh/trae020

Abstract

Background: Melioidosis is caused by Burkholderia pseudomallei. Direct molecular detection from unamplified blood remains insensitive.

Methods: 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.

Results: The QIAamp UCP Pathogen Mini Kit performed best, with a limit of detection of 1.5×102 cfu/ml.

Conclusions: It is planned to use the QIAamp UCP Pathogen Mini Kit to do a larger study on blood collected from patients with melioidosis.

Keywords: Burkholderia pseudomallei; DNA; PCR blood; extraction; melioidosis.