Table 2.
HIV DNA real time PCR assays applied in HAND studies.
| Reference | Target | Sample | Clinical Data | Study Outcome | Advantage/Disadvantage |
|---|---|---|---|---|---|
| [73] | gag | PBMC | HAD patients on HAART 20–39 years and 50 years and above |
Quantities of HIV DNA correlated with HAD HIV was higher in activated monocytes than CD14− in two patients |
Very sensitive Quantitect Sybr Green PCR assay with detection limit of 1–3 copies and applicable suppressed patients and those with low cell PBMC or cell subset counts |
| [78] | gag | PBMC, CD14+, CD14− | 15 HAART-naïve HAD patients and 15 Non demented patients Average age, 34.1 |
HIV DNA in PBMC was significantly higher in HAD non demented patients HIV DNA in five HAD patients was higher in CD14/CD16 than CD4 |
Very sensitive Quantitect Sybr Green PCR assay with detection limit of 1–3 copies and can be applied to suppressed patients and samples with low cell count |
| [31] | pol | PBMC | HIV Subtype B chronic patients on cART for six months CD4+ < 350/mL Median Age 56, 74 |
HIV DNA was associated with HAD and not Non demented forms of HAND |
Very sensitive assay with one copy as LOD, hence suitable for supressed HAND patients PCR has a wide dynamic range from three copies to 300,000 copies High efficiencies for both HIV DNA and β-actin normaliser (93.7% and 86.6% respectively) -Assay restricted to detecting Subtype B |
| [42] | LTR-gag | CD14− and CD14+ | ART naïve participants (median age, 32) with 17 impaired and 19 unimpaired Patients had higher HIV DNA in lymphocytes than monocytes |
Strong association between HIV DNA levels in lymphocytes and HAND | The target region for amplification is highly conserved and the Universal PCR assay has been customised for subtype G and CRF02_AG. However, performance of the assays not detailed |
| [79] | gag | CD14+ | 12 HAART treated HAD and 15 non HAD Infected with HIV-1 CRF01_AE Median Age, 32 |
Baseline monocyte HIV DNA correlated with HIV DNA and 48 weeks after HAART. Monocyte HIV DNA level was below detection limit in all non-HAD patients after 48 weeks |
Detection limit of the assay was 10 copies/106 cells which is relatively high. |
| [11] | gag | CD14+PBMC | Treatment naïve with three clinical categories of HAND (ANI, MND, HAD) CD4+ <350/mL –CRF AE_01Mean Age, 35 |
No correlation between PBMC HIV DNA Before CD14+ enrichment, CD14+ was associated with HAND(ANI, MND, HAD) –CD14+ HIV DNA was associated with glial dysfunction, neuronal injury and CSF immune activation (neopterin) |
Very sensitive assay with a limit of detection of one copy which is suitable for HAND suppressed patients |
| [51] | pol | PBMC | 22–40 years and 50–71 years ART-supressed HIV subjects | Higher HIV DNA levels were associated with more severe neurocognitive impairment in older patients No association between HIV DNA and HAND in young adults. |
Very sensitive droplet digital PCR with Limit of Detection (LOD) of one copy and more precise than qPCR due insensitivity to primer and probe mismatches absolute quantification, no external standards required |