Quantification of HIV-1 proviral DNA and analysis of genomic diversity by polymerase chain reaction and temperature gradient gel electrophoresis

Wieland, Ulrike; Suhr, Helge; Salzberger, Bernd; Eggers, Hans J.; Braun, Rüdiger W.; Kühn, Joachim

doi:10.1016/0166-0934(95)01977-4

Cited by 7 publications

(1 citation statement)

References 46 publications

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“…Real time PCR has been used to detect single copies of different viruses from body fluids and tissues (Mackay et al, 2002). Many investigators have described assays capable of detecting a single copy of HIV-1 proviral DNA (Boni et al, 1993; Boni et al, 2004; Bush et al, 1992; McClure et al, 2000; Nuovo, 2000; Saha et al, 2001; Simmonds et al, 1990; Spann et al, 1991; Wieland et al, 1996; Yourno, 1993; Yourno and Conroy, 1992; Zazzi et al, 1992; Zazzi et al, 1993) or HIV RNA (Fischer et al, 2008; Kaiser et al, 2007; Kumar et al, 2007; Palmer et al, 2003) from clinical samples, including peripheral blood mononuclear cells, dried blood spots, and brain tissue. Replicate sampling has been used in single copy assays (Palmer et al, 2003), but can also be used to improve the sensitivity of assays with higher limits of detection.…”

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confidence: 99%

Modification of the Abbott RealTime assay for detection of HIV-1 plasma RNA viral loads less than one copy per milliliter

Yukl

Fujimoto

et al. 2011

Journal of Virological Methods

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Abstract/Summary Although commercial tests are approved for detection of HIV-1 plasma viral loads ≥20 copies per milliliter (ml), only one specialized research assay has been reported to detect plasma viral loads as low as 1 copy/ml. This manuscript describes a method of concentrating HIV-1 virions from up to 30ml of plasma, which can be combined with a commercial viral load test to create a widely-available, reproducible assay for quantifying plasma HIV RNA levels less than 1 copy/ml. Using this pre-analytically modified assay, samples with a known level of 0.5 copy/ml were detected in 8 of 12 replicates (mean 0.47 copy/ml; 95% confidence interval (CI) 0.14-0.81 copy/ml) and samples with a known level of 1.0 copy/ml were detected in 13 of 13 replicates (mean 1.96 copy/ml; 95% CI 1.42-2.50 copy/ml). By concentrating virus from 30ml of plasma, HIV RNA could be measured in 16 of 19 samples (84%) from 12 of 12 subjects (mean 2.77 copy/ml; 95% CI 0.86-4.68 copy/ml). The measured viral load correlated inversely (r= −0.78; p=0.028) with the total duration of viral suppression (viral load<40 copies/ml).

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