Quantitation of viral DNA by real-time PCR applying duplex amplification, internal standardization, and two-color fluorescence detection.
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 10(2) to at least 5 x 10(6) 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.
Gruber
F
F
Hyland-Immuno Division, Biomedical Research Center, Baxter, A-2304 Orth-Donau, Austria.
Falkner
F G
FG
Dorner
F
F
Hämmerle
T
T
D000465:Algorithms; D015342:DNA Probes; D004279:DNA, Viral / Q000032:analysis*; D005453:Fluorescence; D016732:Parvovirus B19, Human / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D012680:Sensitivity and Specificity
D015342:DNA Probes; D004279:DNA, Viral