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Einsatzrelevanz der PCR als diagnostisches Verfahren

Mission oriented diagnostic real-time PCR

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Summary

In out of area military missions soldiers are potentially exposed to bacteria that are endemic in tropical areas and can be used as biological agents. It can be difficult to culture these bacteria due to sample contamination, low number of bacteria or pretreatment with antibiotics. Commercial biochemical identification systems are not optimized for these agents which can result in misidentification. Immunological assays are often not commercially available or not specific. Real-time PCR assays are very specific and sensitive and can shorten the time required to establish a diagnosis markedly. Therefore, real-time PCRs for the identification of Bacillus anthracis, Brucella spp., Burkholderia mallei und Burkholderia pseudomallei, Francisella tularensis und Yersinia pestis have been developed. PCR results can be false negative due to inadequate clinical samples, low number of bacteria in samples, DNA degradation, inhibitory substances and inappropriate DNA preparation. Hence, it is crucial to cultivate the organisms as a prerequisite for adequate antibiotic therapy and typing of the agent. In a bioterrorist scenario samples have to be treated according to rules applied in forensic medicine and documentation has to be flawless.

Zusammenfassung

Bei militärischen Auslandseinsätzen können Soldaten durch Bakterien gefährdet sein, die in den Tropen endemisch sind und gleichzeitig potentielle B-Kampfstoffe darstellen. Die Anzucht dieser Keime aus klinischen Proben gestaltet sich wegen Kontamination, geringer Keimzahl oder antibiotischer Vorbehandlung häufig sehr problematisch. Kommerzielle biochemische Identifizierungsmethoden sind für diese Keime nicht optimiert und daher unzuverlässig. Immunologische Nachweisverfahren sind häufig kommerziell nicht verfügbar oder nicht ausreichend spezifisch. Real-time PCR-Methoden sind im Gegensatz hierzu sehr spezifisch und sensitiv und können die Diagnostik wesentlich beschleunigen. Real-time PCRs für den Nachweis von Bacillus anthracis, Brucella spp., Burkholderia mallei und Burkholderia pseudomallei, Francisella tularensis und Yersinia pestis sind daher entwickelt worden. Die Ergebnisse der PCR-Diagnostik können durch ungeeignete Primärproben, geringe Erregermenge in der Probe, Degradation der DNA, inhibitorische Substanzen und unzureichende DNA-Präparation zu falsch negativen Ergebnissen führen. Die Anzucht des Erregers ist immer zu versuchen, weil sie gezielte antibiotische Therapie und eine weitere Typisierung des Erregers erlaubt. Bei möglicherweise bio-terroristischem Hintergrund ist mit Probenmaterial nach den Regeln der forensischen Medizin umzugehen und die Dokumentation muss lückenlos sein.

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Authors and Affiliations

  1. Militärspital Innsbruck, Innsbruck, Austria

    Herbert Tomaso

  2. Institut für Mikrobiologie der Bundeswehr, München, Germany

    Herbert Tomaso, Holger C. Scholz, Wolf D. Splettstoesser & Martin Pfeffer

  3. Bundeswehrzentralkrankenhaus Koblenz, Koblenz, Germany

    Sascha Al Dahouk

  4. Friedrich-Loeffler-Institut, Jena, Germany

    Heinrich Neubauer

  5. Institut für Medizinische Mikrobiologie, Jena, Germany

    Eberhard Straube

Authors
  1. Herbert Tomaso
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  2. Holger C. Scholz
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  3. Sascha Al Dahouk
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  4. Wolf D. Splettstoesser
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  5. Heinrich Neubauer
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  6. Martin Pfeffer
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  7. Eberhard Straube
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Correspondence to Herbert Tomaso.

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Tomaso, H., Scholz, H., Al Dahouk, S. et al. Einsatzrelevanz der PCR als diagnostisches Verfahren. Wien Klin Wochenschr 119 (Suppl 3), 26–32 (2007). https://doi.org/10.1007/s00508-007-0858-4

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  • DOI: https://doi.org/10.1007/s00508-007-0858-4

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