Comparison of the utility of five commercial kits for extraction of DNA from Aspergillus fumigatus spores

• Authors: Urszula Nawrot , Katarzyna Wlodarczyk , Magdalena Wrobel , Anita Wasik , Tadeusz Dobosz
• Languages of publication: EN

Abstracts

EN

The aim of this study was to compare the efficiency of DNA extraction from water as well as from blood samples spiked with A. fumigatus spores, using selected commercial kits. Extraction of DNA according to manufacturer's protocols was preceded by blood cells lysis and disruption of fungal cells by enzymatic digestion or bead beating. The efficiency of DNA extraction was measured by PCR using Aspergillus-specific primers and SYBR Green I dye or TaqMan probes targeting 28S rRNA gene. All methods allowed the detection of Aspergillus at the lowest tested density of water suspensions of spores (101 cells/ml). The highest DNA yield was obtained using the ZR Fungal/Bacterial DNA kit, YeastStar Genomic DNA kit, and QIAamp DNA Mini kit with mechanical cell disruption. The ZR Fungal/Bacterial DNA and YeastStar kits showed the highest sensitivity in examination of blood samples spiked with Aspergillus (100 % for the detection of 102 spores and 75 % for 101 spores). Recently, the enzymatic method ceased to be recommended for examination of blood samples for Aspergillus, thus ZR Fungal/Bacterial DNA kit and QIAamp DNA Mini kit with mechanical cell disruption could be used for extraction of Aspergillus DNA from clinical samples.

Keywords

EN

DNA extraction; aspergillosis; Aspergillus

• Publisher: Polish Biochemical Society
• Journal: Acta Biochimica Polonica
• Year: 2010
• Volume: 57
• Issue: 4
• Pages: 567-571

Dates

published

2010

received

2010-06-24

revised

2010-08-25

accepted

2010-10-25

(unknown)

2010-10-27

Contributors

author

Urszula Nawrot

• Department of Microbiology, Wroclaw Medical University, Wrocław, Poland

author

Katarzyna Wlodarczyk

• Department of Microbiology, Wroclaw Medical University, Wrocław, Poland

author

Magdalena Wrobel

• Department of Microbiology, Wroclaw Medical University, Wrocław, Poland

author

Anita Wasik

• Department of Microbiology, Wroclaw Medical University, Wrocław, Poland

author

Tadeusz Dobosz

• Molecular Techniques Unit, Wroclaw Medical University, Wrocław, Poland

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