scholarly journals A novel TaqMan real-time PCR assay to estimate ex vivo human immunodeficiency virus type 1 fitness in the era of multi-target (pol and env) antiretroviral therapy

2003 ◽  
Vol 84 (8) ◽  
pp. 2217-2228 ◽  
Author(s):  
Jan Weber ◽  
Hector R. Rangel ◽  
Bikram Chakraborty ◽  
Mahlet Tadele ◽  
Miguel A. Martinez ◽  
...  
Keyword(s):  
Human Immunodeficiency Virus ◽  
Antiretroviral Therapy ◽  
Real Time ◽  
Human Immunodeficiency Virus Type ◽  
Virus Type ◽  
Real Time Pcr ◽  
Ex Vivo ◽  
Pcr Assay ◽  
Immunodeficiency Virus

scholarly journals Quantification of Human Immunodeficiency Virus Type 1 Proviral Load by a TaqMan Real-Time PCR Assay

2001 ◽  
Vol 39 (4) ◽  
pp. 1303-1310 ◽  
Author(s):  
N. Desire ◽  
A. Dehee ◽  
V. Schneider ◽  
C. Jacomet ◽  
C. Goujon ◽  
...  
Keyword(s):  
Human Immunodeficiency Virus ◽  
Real Time ◽  
Human Immunodeficiency Virus Type ◽  
Virus Type ◽  
Real Time Pcr ◽  
Proviral Load ◽  
Pcr Assay ◽  
Immunodeficiency Virus

Evaluation of the COBAS Ampliprep/COBAS TaqMan System for Quantification of Human Immunodeficiency Virus type 1 (HIV-1) RNA by Real-time PCR

2008 ◽  
Vol 82 (1) ◽  
pp. 20-25
Author(s):  
Katsuji TERUYA ◽  
Shinichi OKA ◽  
Katsuyuki FUKUTAKE ◽  
Kagehiro AMANO ◽  
Shigeyuki FURUTANI ◽  
...  
Keyword(s):  
Human Immunodeficiency Virus ◽  
Real Time ◽  
Human Immunodeficiency Virus Type ◽  
Virus Type ◽  
Real Time Pcr ◽  
Cobas Taqman ◽  
Immunodeficiency Virus ◽  
Hiv 1

scholarly journals Development of a New Methodology for Screening of Human Immunodeficiency Virus Type 1 Microbicides Based on Real-Time PCR Quantification

2006 ◽  
Vol 51 (2) ◽  
pp. 638-644 ◽  
Author(s):  
Renato S. Aguiar ◽  
Luciana J. Costa ◽  
Helena S. Pereira ◽  
Rodrigo M. Brindeiro ◽  
Amilcar Tanuri
Keyword(s):  
Human Immunodeficiency Virus ◽  
Reverse Transcriptase ◽  
Real Time ◽  
Human Immunodeficiency Virus Type ◽  
Virus Type ◽  
Real Time Pcr ◽  
Immunodeficiency Virus ◽  
The Impact ◽  
Hiv 1

ABSTRACT Potential topical retrovirucides or vaginal microbicides against human immunodeficiency virus type 1 (HIV-1) include nonnucleoside reverse transcriptase inhibitors (NNRTIs). To be successful, such agents have to be highly active against cell-free virions. In the present study, we developed a new real-time PCR-based assay to measure the natural endogenous reverse transcription (NERT) activity directly on intact HIV-1 particles in the presence of reverse transcriptase (RT) inhibitors. We further evaluated the permeability to nevirapine (NVP) and efavirenz (EFV) and their retention within nascent viral particles. We also demonstrated the NVP and EFV inhibitory effects on NERT activity and the impact of resistance mutations measured directly by this new strategy. Furthermore, the results showed a clear correlation between NERT activity and classical infectivity assays. The 50% inhibitory concentrations (IC50s) of NVP and EFV were demonstrated to be up to 100-fold higher for cell-free than for cell-associated virions, suggesting that cell-free virions are less permeable to these drugs. Our results suggest that NVP and EFV penetrate both the envelope and the capsid of HIV-1 particles and readily inactivate cell-free virions. However, the characteristics of these NNRTIs, such as lower permeability and lower retention during washing procedures, in cell-free virions reduce their efficacies as microbicides. Here, we demonstrate the usefulness of the NERT real-time PCR as an assay for screening novel antiretroviral compounds with unique mechanisms of action.

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