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Journal of Clinical Microbiology, April 1999, p. 1200-1202, Vol. 37, No. 4
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Contaminations Occurring in Fungal PCR Assays

Juergen Loeffler,1,* Holger Hebart,1 Ralf Bialek,2 Lars Hagmeyer,1 Diethard Schmidt,1 Francois-Prâseth Serey,1 Matthias Hartmann,1 Jan Eucker,3 and Hermann Einsele1

Universität Tuebingen, Medizinische Klinik und Poliklinik, Abteilung II,1 and Institut für Tropenmedizin,2 72076 Tuebingen, and Universitätsklinikum Charité, 10117 Berlin,3 Germany

Received 31 August 1998/Returned for modification 2 November 1998/Accepted 18 December 1998

Successful in vitro amplification of fungal DNA in clinical specimens has been reported recently. In a collaboration among five European centers, the frequency and risk of contamination due to airborne spore inoculation or carryover contamination in fungal PCR were analyzed. The identities of all contaminants were specified by cycle sequencing and GenBank analysis. Twelve of 150 PCR assays that together included over 2,800 samples were found to be contaminated (3.3% of the negative controls were contaminated during the DNA extraction, and 4.7% of the PCR mixtures were contaminated during the amplification process). Contaminants were specified as Aspergillus fumigatus, Saccharomyces cerevisiae, and Acremonium spp. Further analysis showed that commercially available products like zymolyase powder or 10× PCR buffer may contain fungal DNA. In conclusion, the risk of contamination is not higher in fungal PCR assays than in other diagnostic PCR-based assays if general precautions are taken.

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

Journal of Clinical Microbiology, April 1999, p. 1200-1202, Vol. 37, No. 4
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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