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. 2012 Dec 5;7(12):e50859. doi: 10.1371/journal.pone.0050859

Figure 2. Sensitivity and specificity of the SG-PERT assay.

Figure 2

(A) Melting curves of PCR products obtained by SG-PERT assay on the LightCycler® 480 when using 1011 or 104 pU recombinant HIV-1 RT or nuclease-free water (non-template control = NTC) as input for the assay, as indicated. (B, D) Amplification curves of indicated amount of (B) recombinant HIV-1 RT (pU), (D) replication competent HIV-1 (NL4-3 strain) (ng p24/mL) or nuclease-free water (NTC) obtained by SG-PERT on the LightCycler® 480. (C,E) Relation between input of (C) recombinant HIV-1 RT, (E) replication competent HIV-1 (HIV-1), HIV-1 based lentiviral vectors (HIV-1 based vector) or Moloney Murine Leukemia-based retroviral vectors (MoMLV) and obtained cycle of quantification (Cq) values by SG-PERT on the LightCycler® 480. Viral titers in the undiluted samples in (E) (value of “0” on x-axis) were 3,100 ng p24/mL for the replication competent HIV-1 virus, 1.12×107 transducing units/mL (TU/mL) for the HIV-1 based viral vector and 4.9×105 TU/mL for the MoMLV-based vector. Only input levels within linear range of the assay were included for correlation analysis.