Quantification of Total HIV DNA

Total HIV DNA was quantified from PBMC at the enrollment time point for all patients using an in-house, real-time quantitative PCR assay as previously described [31 (link)]. Briefly, total nucleic acid was extracted from PBMCs on the Qiasymphony platform using the DSP virus/pathogen mini kit (Qiagen) according to the manufacturer’s protocol. An ABI PRISM 7500 real-time PCR instrument with Invitrogen RT-PCR reagents was used for amplification and detection of HIV-1 DNA. The HIV DNA copies were calculated as HIV-1 copies per 10^6 haploid pyruvate dehydrogenase (PDH) copies. Results are reported as copies of HIV per million cells [59 (link)].