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Journal of Clinical Microbiology
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Mycology

Contaminations Occurring in Fungal PCR Assays

Juergen Loeffler, Holger Hebart, Ralf Bialek, Lars Hagmeyer, Diethard Schmidt, Francois-Prâseth Serey, Matthias Hartmann, Jan Eucker, Hermann Einsele
Juergen Loeffler
Universität Tuebingen, Medizinische Klinik und Poliklinik, Abteilung II, and
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Holger Hebart
Universität Tuebingen, Medizinische Klinik und Poliklinik, Abteilung II, and
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Ralf Bialek
Institut für Tropenmedizin,72076 Tuebingen, and
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Lars Hagmeyer
Universität Tuebingen, Medizinische Klinik und Poliklinik, Abteilung II, and
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Diethard Schmidt
Universität Tuebingen, Medizinische Klinik und Poliklinik, Abteilung II, and
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Francois-Prâseth Serey
Universität Tuebingen, Medizinische Klinik und Poliklinik, Abteilung II, and
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Matthias Hartmann
Universität Tuebingen, Medizinische Klinik und Poliklinik, Abteilung II, and
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Jan Eucker
Universitätsklinikum Charité, 10117 Berlin, Germany
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Hermann Einsele
Universität Tuebingen, Medizinische Klinik und Poliklinik, Abteilung II, and
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DOI: 10.1128/JCM.37.4.1200-1202.1999
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ABSTRACT

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 asAspergillus 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.

  • Copyright © 1999 American Society for Microbiology
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Contaminations Occurring in Fungal PCR Assays
Juergen Loeffler, Holger Hebart, Ralf Bialek, Lars Hagmeyer, Diethard Schmidt, Francois-Prâseth Serey, Matthias Hartmann, Jan Eucker, Hermann Einsele
Journal of Clinical Microbiology Apr 1999, 37 (4) 1200-1202; DOI: 10.1128/JCM.37.4.1200-1202.1999

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Contaminations Occurring in Fungal PCR Assays
Juergen Loeffler, Holger Hebart, Ralf Bialek, Lars Hagmeyer, Diethard Schmidt, Francois-Prâseth Serey, Matthias Hartmann, Jan Eucker, Hermann Einsele
Journal of Clinical Microbiology Apr 1999, 37 (4) 1200-1202; DOI: 10.1128/JCM.37.4.1200-1202.1999
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KEYWORDS

fungi
polymerase chain reaction

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