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Applied and Environmental Microbiology, June 2001, p. 2837-2839, Vol. 67, No. 6
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.6.2837-2839.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Quantitation of Viral DNA by Real-Time PCR Applying Duplex Amplification, Internal Standardization, and Two-Color Fluorescence Detection

Franz Gruber, Falko G. Falkner, Friedrich Dorner, and Thomas Hämmerle*

Hyland-Immuno Division, Biomedical Research Center, Baxter, A-2304 Orth-Donau, Austria

Received 6 September 2000/Accepted 19 March 2001

A real-time PCR method was developed to quantitate viral DNA that includes duplex amplification, internal standardization, and two-color fluorescence detection without the need to generate an external standardization curve. Applied to human parvovirus B19 DNA, the linear range was from 102 to at least 5 × 106 copies per ml of sample. The coefficient of variation was 0.29 using a run control of 2,876 copies per ml. The method reduces the risk of false-negative results, yields high precision, and is applicable for other DNA targets.

* Corresponding author. Mailing address: Baxter, Hyland-Immuno Division, Biomedical Research Center, Uferstr. 15, A-2304 Orth-Donau, Austria. Phone: 43-1-20100-4306. Fax: 43-1-20100-4000. E-mail: haemmet{at}baxter.com.

Applied and Environmental Microbiology, June 2001, p. 2837-2839, Vol. 67, No. 6
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.6.2837-2839.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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