PMID: 9179766Jun 1, 1997Paper

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

Journal of Medical Virology
F KashanchiM R Sadaie

Abstract

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) and subsequently assayed by both in situ RNA hybridization and a reverse transcriptase polymerase chain reaction (RT-PCR). The sensitivity of RT-PCR was estimated to be equivalent to 1 x 10(-16) grams (0.1 fg) or approximately 64 copies of the input standard viral RNA per reaction. The present study takes advantage of the ability of NaB to introduce changes in chromatin structure of latently infected cells, leading to increased HIV gene expression. Human ACH-2 and U1 cell lines were used as representatives of T-lymphocytic and monocytoid cells harboring latent inducible proviruses. HIV gene expression was readily detected when these cells were treated with NaB. Viral gag RNA was detected by both in situ and RT-PCR assays. When peripheral blood mononuclear cells (PBMCs) from acquired immunodeficiency syndrome (AIDS) patients, who were all negative for in situ hybridization and serum/plasma p24 assays, were used for detection of viral gene expression, four categories with distinct patterns of induction were observed. ...Continue Reading

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Citations

Aug 9, 2006·Retrovirology·Emmanuel AgbottahFatah Kashanchi
Sep 26, 2015·Clinical Epigenetics·Amit KumarGeorges Herbein
Feb 24, 2000·Journal of Medical Virology·Y L LinH S Liu
Nov 9, 2004·The Journal of Biological Chemistry·Emmanuel AgbottahFatah Kashanchi
Mar 7, 2009·The Journal of Immunology : Official Journal of the American Association of Immunologists·Kenichi ImaiTakashi Okamoto

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