Author:

Juergen Loeffler, Kathrin Schmidt, Holger Hebart, Ulrike Schumacher, and Hermann Einsele

Date: 1 September 2002

Abstract:

A fully automated assay was established for the extraction of DNA from clinically important fungi by usingthe MagNA Pure LC instrument. The test was evaluated by DNA isolation from 23 species of yeast andfilamentous 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. Thedetection limit of the MagNA Pure LC assay of 10 CFU corresponded to the sensitivity when DNA was extractedmanually; in 9 of 28 runs, we could achieve a higher sensitivity of 1 CFU/ml blood, which was found to besignificant (p < 0.004). DNA from all fungal species analyzed could be extracted and amplified by real-timePCR. Negative controls from all MagNA Pure isolations remained negative. Sixty-three clinical samplesshowed 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.

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