Quantitative RNA and DNA gene amplification can rapidly monitor HIV infection and antiviral activity in cell cultures.
We have developed a quantitative gene amplification procedure to assess the replication of human immunodeficiency virus (HIV) in cell cultures and evaluate the effect of drugs on viral replication. Increases in HIV gag RNA and DNA in phytohemagglutinin-stimulated normal peri-pheral blood mononuclear cells (PBMC) infected with HIV at very low multiplicity of infection paralleled the production of HIV p24 antigen in culture supernatants. Quantitative gene amplification was able to monitor the accumulation of viral nucleic acids in control cultures and demonstrate the effect of various concentrations of azidothymidine (AZT) on the replication of both AZT-sensitive and -resistant strains of HIV. The sensitivity of patient-derived virus strains to AZT could also be successfully measured by these procedures. The results of our studies suggest that quantitative measurement of HIV gag RNA and DNA can be used to monitor the kinetics of viral replication, antiviral activity, viral drug resistance, and mechanism of drug action.
Winters
M A
MA
Center for AIDS Research, Stanford University Medical Center, California 94305-5107.
Holodniy
M
M
Katzenstein
D A
DA
Merigan
T C
TC
D002478:Cells, Cultured; D004279:DNA, Viral / Q000032:analysis*; D015750:Genes, gag; D006678:HIV / Q000187:drug effects / Q000235:genetics* / Q000302:isolation & purification / Q000502:physiology; D016655:HIV Core Protein p24 / Q000032:analysis; D015658:HIV Infections / Q000382:microbiology*; D006679:HIV Seropositivity / Q000382:microbiology; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000382:microbiology*; D008213:Lymphocyte Activation; D012367:RNA, Viral / Q000032:analysis*; D014779:Virus Replication / Q000187:drug effects*; D015215:Zidovudine / Q000494:pharmacology*
D004279:DNA, Viral; D016655:HIV Core Protein p24; D012367:RNA, Viral; D015215:Zidovudine