Table 2. Detection of integrated HIV-1 genome in epithelial cells by qPCR.
Cell line | R5-YU2 (MOI = 7.5) | X4-LAI (MOI = 10) | X4-LAI (MOI = 140) |
FaDu | ND | Not included | ND |
TR146 | ND | ND | ND |
A431 | ND | ND | ND |
C8166a | n/a | + (Ct = 33) | + (Ct = 33) |
NP2-R5b | + (Ct = 29) | n/a | n/a |
C8166 cells express CXCR4 and were used for X4 viral infections only.
NP2-R5 cells express CCR5 and were used for R5 viral infections only.
+, Integrated HIV-1 product detected (cycle threshold detection in brackets).
ND, Integrated HIV-1 product not detected.
Real-time PCR assay to detect integrated HIV-DNA using HIV-1 LTR and human Alu-specific primers and a U5 specific probe [92]. TR146, FaDu, A431 and PM-1 (control) cells were exposed to YU2 (R5) or LAI (X4) virus for 48 h at 37°C. DNA was extracted, digested with DpnI to degrade any plasmid DNA contaminant and analysed by real-time PCR.