Comparison of four commercial DNA extraction kits for the recovery of Bacillus spp. spore DNA from spiked powder samples

Bacillus spp. include human pathogens such as Bacillus anthracis, the causative agent of anthrax and a biothreat agent. Bacillus spp. form spores that are physically highly resistant and may remain active over sample handling. We tested four commercial DNA extraction kits (QIAamp DNA Mini Kit, RTP P...

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Bibliographic Details
Published in:Journal of microbiological methods 2016-09, Vol.128, p.69-73
Main Authors: Mölsä, Markos, Kalin-Mänttäri, Laura, Tonteri, Elina, Hemmilä, Heidi, Nikkari, Simo
Format: Article
Language:English
Subjects:
Bacillus anthracis
Bacillus spp
Bacillus thuringiensis
Bacillus thuringiensis - isolation & purification
Biothreat
DNA extraction
DNA, Bacterial - isolation & purification
Limit of Detection
Powder
Powders - chemistry
Reagent Kits, Diagnostic
Solanum tuberosum
Spore
Spore inactivation
Spores, Bacterial - isolation & purification
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Summary:Bacillus spp. include human pathogens such as Bacillus anthracis, the causative agent of anthrax and a biothreat agent. Bacillus spp. form spores that are physically highly resistant and may remain active over sample handling. We tested four commercial DNA extraction kits (QIAamp DNA Mini Kit, RTP Pathogen Kit, ZR Fungal/Bacterial DNA MiniPrep, and genesig Easy DNA/RNA Extraction kit) for sample inactivation and DNA recovery from two powders (icing sugar and potato flour) spiked with Bacillus thuringiensis spores. The DNA was analysed using a B. thuringiensis-specific real-time PCR assay. The detection limit was 3×101CFU of spiked B. thuringiensis spores with the QIAamp DNA Mini, RTP Pathogen, and genesig Easy DNA/RNA Extraction kits, and 3×103CFU with the ZR Fungal/Bacterial DNA MiniPrep kit. The results showed that manual extraction kits are effective and safe for fast and easy DNA extraction from powder samples even in field conditions. Adding a DNA filtration step to the extraction protocol ensures the removal of Bacillus spp. spores from DNA samples without affecting sensitivity. •Bacillus spp. spores may remain active over sample handling.•Performance of manual DNA extraction kits were tested for sample inactivation.•DNA samples were incubated on LB plates to evaluate the inactivation efficiency.•Adding a DNA filtration step ensured the removal of viable bacteria and spores.•Manual extraction kits are effective for fast DNA extraction in field conditions.
ISSN:0167-7012
1872-8359
DOI:10.1016/j.mimet.2016.07.013