title: Comparison of nucleic acid extraction platforms for detection of select biothreat agents for use in clinical resource limited settings.
abstract: High-quality nucleic acids are critical for optimal PCR-based diagnostics and pathogen detection. Rapid sample processing time is important for the earliest administration of therapeutic and containment measures, especially in the case of biothreat agents. In this context, we compared the Fujifilm QuickGene-Mini80 to Qiagen's QIAamp Mini Purification kits for extraction of DNA and RNA for potential use in austere settings. Qiagen (QIAamp) column-based extraction is the currently recommended purification platform by United States Army Medical Research Institute for Infectious Diseases for both DNA and RNA extraction. However, this sample processing system requires dedicated laboratory equipment including a centrifuge. In this study, we investigated the QuickGene-Mini80, which does not require centrifugation, as a suitable platform for nucleic acid extraction for use in resource-limited locations. Quality of the sample extraction was evaluated using pathogen-specific, real-time PCR assays for nucleic acids extracted from viable and γ-irradiated Bacillus anthracis, Yersinia pestis, vaccinia virus, Venezuelan equine encephalitis virus, or B. anthracis spores in buffer or human whole blood. QuickGene-Mini80 and QIAamp performed similarly for DNA extraction regardless of organism viability. It was noteworthy that γ-irradiation did not have a significant impact on real-time PCR for organism detection. Comparison with QIAamp showed a less than adequate performance of the Fujifilm instrument for RNA extraction. However, QuickGene-Mini80 remains a viable alternative to QIAamp for DNA extraction for use in remote settings due to extraction quality, time efficiency, reduced instrument requirements, and ease of use.
authors: Diagnostic Systems Division, US Army Medical Research Institute of Infectious Diseases, Fort Detrick, MD, USA.
authors: Michelle A
authors: MA
authors: Shipley
authors: Jeffrey W
authors: JW
authors: Koehler
authors: David A
authors: DA
authors: Kulesh
authors: Timothy D
authors: TD
authors: Minogue
mesh_terms: D001419:Bacteria / Q000235:genetics / Q000302:isolation & purification*; D054045:Biological Warfare Agents; D004247:DNA / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D008828:Microbiological Techniques / Q000379:methods*; D008967:Molecular Biology / Q000379:methods*; D012313:RNA / Q000235:genetics / Q000302:isolation & purification*; D011933:Reagent Kits, Diagnostic; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D014481:United States; D014780:Viruses / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D054045:Biological Warfare Agents; D011933:Reagent Kits, Diagnostic; D012313:RNA; D004247:DNA