Rapid and sensitive detection of cell-associated HIV-1 in latently infected cell lines and in patient cells using sodium-n-butyrate induction and RT-PCR

摘要: To develop a rapid and sensitive means of detecting cell-associated human immunodeficiency virus (HIV), donor cells from HIV seropositive patients were treated with the potent viral activator sodium-n-butyrate (NaB) subsequently assayed by both in situ RNA hybridization reverse transcriptase polymerase chain reaction (RT-PCR). The sensitivity RT-PCR was estimated to be equivalent 1 x 10(-16) grams (0.1 fg) or approximately 64 copies input standard per reaction. present study takes advantage ability NaB introduce changes chromatin structure latently infected cells, leading increased gene expression. Human ACH-2 U1 cell lines used as representatives T-lymphocytic monocytoid harboring latent inducible proviruses. expression readily detected when these NaB. Viral gag assays. When peripheral blood mononuclear (PBMCs) acquired syndrome (AIDS) patients, who all negative for serum/plasma p24 assays, detection expression, four categories distinct patterns induction observed. first set showed HIV-positive PBMCs without any added NaB, suppression PHA. second samples alone. third could induced PHA, but not fourth required PHA Our results suggest that direct treatment activators may useful increasing intended RNAs. This approach confirm true status infection are RNAs below threshold detection. Moreover, this method identify presence proviral genomes possibly reflecting rate load vivo possible mutations brought about long-term co-cultivation assays seronegative donors.