Table 2

Measurement of viral RNA in concentrated lentiviral vector preparations

Ct value


DNA Standard

LTR primers

eGFP primers

WPRE primers


5.0 × 108

15.51 ± 0.01

15.65 ± 0.04

15.45 ± 0.06

5.0 × 107

17.57 ± 0.01

17.79 ± 0.10

17.51 ± 0.01

5.0 × 106

18.97 ± 0.06

19.11 ± 0.08

20.11 ± 0.05

5.0 × 105

22.06 ± 0.17

21.82 ± 0.07

22.59 ± 0.03

5.0 × 104

25.91 ± 0.05

24.06 ± 0.06

26.13 ± 0.04


RNA extracts

CH-eGFP-WS

22.87 ± 0.05

22.5 ± 0.02

22.04 ± 0.03

CH-eGFP-WS

16.15 ± 0.02

16.57 ± 0.03

16.43 ± 0.05


Primer/probe sets annealing to the front (LTR), the centre (GFP) or at the end (WPRE) of the genomic RNA of lentiviral vectors were used to determine to what extent full-length genomic vector RNA is incorporated into lentiviral vector particles. In one-step RT-qPCR assays with the different primer/probe sets comparable threshold cycles (Ct) were detected when amplifying a dilution series of the pCH-eGFP-WS transfer plasmid (DNA standard) or RNA extracts of two representative CH-eGFP-WS vector preparations. Mean Ct values ± standard deviation for 3 amplifications of the same sample are shown.

Geraerts et al. BMC Biotechnology 2006 6:34   doi:10.1186/1472-6750-6-34

Open Data