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Journal of Clinical Microbiology, December 2001, p. 4413-4419, Vol. 39, No. 12
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.12.4413-4419.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

New LightCycler PCR for Rapid and Sensitive Quantification of Parvovirus B19 DNA Guides Therapeutic Decision-Making in Relapsing Infections

Timm C. Harder,1,* Markus Hufnagel,2,dagger Katrin Zahn,1 Karin Beutel,2 Heinz-Josef Schmitt,2,Dagger Uwe Ullmann,1 and Peter Rautenberg1

Department of Medical Microbiology and Virology1 and University Children's Hospital,2 Christian-Albrechts University, Kiel, Germany

Received 27 November 2000/Returned for modification 20 March 2001/Accepted 27 September 2001

Detection of parvovirus B19 DNA offers diagnostic advantages over serology, particularly in persistent infections of immunocompromised patients. A rapid, novel method of B19 DNA detection and quantification is introduced. This method, a quantitative PCR assay, is based on real-time glass capillary thermocycling (LightCycler [LC]) and fluorescence resonance energy transfer (FRET). The PCR assay allowed quantification over a dynamic range of over 7 logs and could quantify as little as 250 B19 genome equivalents (geq) per ml as calculated for plasmid DNA (i.e., theoretically >= 5 geq per assay). Interrater agreement analysis demonstrated equivalence of LC-FRET PCR and conventional nested PCR in the diagnosis of an active B19 infection (kappa coefficient = 0.83). The benefit of the new method was demonstrated in an immunocompromised child with a relapsing infection, who required an attenuation of the immunosuppressive therapy in addition to repeated doses of immunoglobulin to eliminate the virus.

* Corresponding author. Present address: Food and Veterinary Diagnostic Laboratory, Max-Eyth Strasse 5, D-24537 Neumuenster, Germany. Phone: 49 4321 904 766. Fax: 49 4321 766 619. E-mail: timm.harder{at}lvua-sh.de.

dagger Present address: Channing Laboratory, Brigham and Women's Hospital, Department of Medicine, Boston, MA 02115.

Dagger Present address: Center for Preventive Pediatrics, Department of Pediatric Infectious Diseases, University Children's Hospital, Johannes-Gutenberg University, D-55101 Mainz, Germany.

Journal of Clinical Microbiology, December 2001, p. 4413-4419, Vol. 39, No. 12
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.12.4413-4419.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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