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Journal of Clinical Microbiology, April 2006, p. 1283-1287, Vol. 44, No. 4
0095-1137/06/$08.00+0     doi:10.1128/JCM.44.4.1283-1287.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Follow-Up on Diagnostic Proficiency of Laboratories Equipped To Perform Orthopoxvirus Detection and Quantification by PCR: the Second International External Quality Assurance Study{dagger}

Matthias Niedrig,1 Hermann Meyer,2 Marcus Panning,3 and Christian Drosten3*

Robert Koch Institute, Berlin,1 Bundeswehr Institute of Microbiology, Munich,2 Bernhard Nocht Institute for Tropical Medicine, Hamburg, Germany3

Received 10 October 2005/ Returned for modification 5 December 2005/ Accepted 2 February 2006

Two years after the first external quality assurance study on bioterrorism-relevant viruses, we have conducted a follow-up study on orthopoxvirus detection by PCR. Thirty-three laboratories (27 European, 4 Austral-Asian, and 2 American) participated. Samples contained 0 to 40,000,000 DNA copies of lyophilized monkeypox, cowpox, and vaccinia virus per ml. Laboratories achieved a >80% detection chance above 56,234 copies per ml. Global sensitivity was not significantly improved over that of the first study. Twenty-seven and 9 participants, respectively, were able to genotype and quantify virus. Four of 27 genotyping results were incorrect. Quantification accuracy was significantly better for vaccinia virus than for the other viruses. False-positive results occurred in 22 (11.8%) of all 186 tests on negative samples, but 18 of these were contributed by only five laboratories. Fifty-five percent of laboratories could appropriately detect PCR inhibition. The use of either real-time PCR or commercial diagnostic kits had significant positive influence on laboratory performance.

* Corresponding author. Mailing address: Department of Virology, Bernhard Nocht Institute for Tropical Medicine, Bernhard-Nocht Str. 74, 20359 Hamburg, Germany. Phone: 49-40-42818-490. Fax: 49-40-42818-378. E-mail: drosten{at}bni-hamburg.de.

{dagger} Supplemental material for this article may be found at http://jcm.asm.org/.

Journal of Clinical Microbiology, April 2006, p. 1283-1287, Vol. 44, No. 4
0095-1137/06/$08.00+0     doi:10.1128/JCM.44.4.1283-1287.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Fedele, C. G., Negredo, A., Molero, F., Sanchez-Seco, M. P., Tenorio, A. (2006). Use of Internally Controlled Real-Time Genome Amplification for Detection of Variola Virus and Other Orthopoxviruses Infecting Humans. J. Clin. Microbiol. 44: 4464-4470 [Abstract] [Full Text]  


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