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Journal of Clinical Microbiology, June 2002, p. 2240-2243, Vol. 40, No. 6
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.6.2240-2243.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Automated Extraction of Genomic DNA from Medically Important Yeast Species and Filamentous Fungi by Using the MagNA Pure LC System

Juergen Loeffler,1* Kathrin Schmidt,1 Holger Hebart,1 Ulrike Schumacher,2 and Hermann Einsele1

Medizinische Klinik, Eberhard-Karls-Universität Tuebingen,1 Institut für Medizinische Mikrobiologie, 72076 Tuebingen, Germany2

Received 21 May 2001/ Returned for modification 24 September 2001/ Accepted 4 March 2002

A fully automated assay was established for the extraction of DNA from clinically important fungi by using the MagNA Pure LC instrument. The test was evaluated by DNA isolation from 23 species of yeast and filamentous fungi and by extractions (n = 28) of serially diluted Aspergillus fumigatus conidia (105 to 0 CFU/ml). Additionally, DNA from 67 clinical specimens was extracted and compared to the manual protocol. The detection limit of the MagNA Pure LC assay of 10 CFU corresponded to the sensitivity when DNA was extracted manually; in 9 of 28 runs, we could achieve a higher sensitivity of 1 CFU/ml blood, which was found to be significant (p <= 0.004). DNA from all fungal species analyzed could be extracted and amplified by real-time PCR. Negative controls from all MagNA Pure isolations remained negative. Sixty-three clinical samples showed identical results by both methods, whereas in 4 of 67 samples, discordant results were obtained. Thus, the MagNA Pure LC technique offers a fast protocol for automated DNA isolation from numerous fungi, revealing high sensitivity and purity.

* Corresponding author. Mailing address: Medizinische Klinik, Abt. II, Labor Prof. Dr. med. H. Einsele, Otfried-Mueller-Str. 10, 72076 Tuebingen, Germany. Phone: 49 7071 2987355. Fax: 49 7071 293179. E-mail: juergen.loeffler{at}med.uni-tuebingen.de.

Journal of Clinical Microbiology, June 2002, p. 2240-2243, Vol. 40, No. 6
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.6.2240-2243.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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