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. Author manuscript; available in PMC: 2015 Feb 1.
Published in final edited form as: J Med Primatol. 2013 Nov 22;43(1):31–43. doi: 10.1111/jmp.12088

Figure 3.

Figure 3

Effects of modifications applied to vRNA extraction and isolation. Panel A. Addition of Proteinase K at 1.0 and 2.5 mg / ml to SIVmac239-containing plasma followed by extraction using the Roche High Pure Viral Kit, cDNA synthesis (using 15 μl vRNA template), and qPCR produced significantly higher SIV PVL results compared to untreated plasma specimens (P < 0.0001, P = 0.0001, respectively). Panel B. Viral RNA recovered after a second elution step that was reverse trasnscribed to cDNA followed by qPCR produced significantly higher SIV PVL results compared to using vRNA from the single elution step in the Roche High Pure Viral Kit (P < 0.0001). Panel C. Viremic plasma was subjected to both Proteinase K digestion and a second elution step to purify vRNA. Different volumes of vRNA template from the modified versus unmodified extraction methods were reverse transcribed for cDNA synthesis followed by qPCR. Although not significant at lower volumes of vRNA, addition of 15 μl of vRNA to the RT followed by qPCR resulted in a significantly higher SIV PVL recovery after application of the dual modification extraction procedure than from the non-modified extraction (P< 0.01). Student’s t Test was used to compare means between groups.