|
Replication event | Assay | Method |
|
Virus Attachment | HeLa-CD4-LT4-β-gal cells | HeLa-cell based assay measuring reduction in chemiluminescence of HIV-1-infected cells [16] |
TZM-bl cells |
gp120/CD4 Ab binding inhibition | Cell based HIV-1 neutralization assay |
gp120 : CD4 ELISA | Biochemical assay with soluble CD4 and monoclonal gp120 antibodies [17] |
gp120/CD4/coreceptor | Cell based, temperature sensitive fusion assay |
|
Fusion and Chemokine Coreceptor Interaction | HL2/3 cells + HeLa-CD4-LTR-β-gal cells | Cell based assay measuring reduction in chemiluminesence [16] |
Coreceptor inhibiton | GHOST-cell based assays measuring reduction in virus replication |
Coreceptor typing | PBMC and Macrophage cell-based assays with tropism-specific clinical HIV-1 isolates [18] |
Compound displacement of chemokine ligands | |
Ca++flux | |
|
Reverse Transcription | Homopolymer and heteropolymer RT inhibition | Biochemical assay measuring reduction in dGTP-[P32] incorporation [19] |
E/intermediate/late RT products | PCR amplification |
RNaseH inhibition | Biochemical assay [20] |
RT inhibition assays using enzymes with specific mutations | Biochemical dGTP-[P32] incorporation assay [19] |
|
Nuclear localization | 2 LTR product in cell nucleus | PCR detection |
|
Integration | Provirus in genomic DNA | PCR detection [21] |
Integrase Complementation | Cell based IN-mutant and Vpr-IN transfection [22] |
Integrase inhibition | Biochemical SPA assay [23] |
Integrase negative virus | |
|
Protein Expression | Northern, Western and flow cytometry | Cell based assays with molecular biology endpoints [24, 25] |
Tat, Rev, and Nef inhibition | Biochemical assays [26, 27] |
Cell-based reporter assay for Rev and Tat function | |
Intracellular p24 | CEM-SS cells infected with HIV-1 and lysed to quantitate p24 by ELISA |
LTR-mediated transcriptional activation | |
|
Protease | Intracellular and virion protein processing | Cell based assay with Western analysis [28] |
Polyprotein cleavage | Biochemical FRET assay [29] |
|