Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2005 Apr;43(4):1851–1857. doi: 10.1128/JCM.43.4.1851-1857.2005

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

PMC Copyright notice

FIG. 2. — Quantitative measurement of HIV-1 proviral DNA content. 8E5 cells were diluted with the whole blood of a healthy HIV-1-seronegative person and spotted to generate five spots with decreasing HIV-1 DNA contents and a constant whole-blood background. A spot with only the seronegative blood was used as the negative DBS control. Extraction of DBS in quadruplicate was carried out in a single run, and the extracted DNA samples were assayed in a single real-time run to determine the intrarun variability (see also Table 2). DBS DNA was extracted and eluted in 50 μl of water, and 20 μl of the eluted DNA was used in the duplex assay. (A) Real-time HIV-1 amplification profiles. The HIV-1 copy numbers per ml of DBS (50 μl of whole blood per circle) are 0 (open circles), 494 (squares), 1,482 (solid triangles), 4,444 (crosses), 13,333 (open triangles), and 40,000 (solid circles). The threshold (cutoff) is 0.354. (B) Real-time RNase P amplification profiles to determine the human chromosomal DNA background of the same dried blood spots as in (A). The threshold is 0.034. (C) HIV-1 content quantification in DBS panel based on threshold cycles (C_Ts) or normalized threshold cycles (dC_Ts). The C_Ts (HIV LTR [diamonds] and RNase P [squares]) were plotted against their respective proviral copy numbers. Each datum was derived from the average of four determinations. Vertical bar: one standard deviation of C_T or dC_T. The dC_T (triangle) is the difference between the HIV C_T and its corresponding RNase P C_T.

FIG. 2. — Quantitative measurement of HIV-1 proviral DNA content. 8E5 cells were diluted with the whole blood of a healthy HIV-1-seronegative person and spotted to generate five spots with decreasing HIV-1 DNA contents and a constant whole-blood background. A spot with only the seronegative blood was used as the negative DBS control. Extraction of DBS in quadruplicate was carried out in a single run, and the extracted DNA samples were assayed in a single real-time run to determine the intrarun variability (see also Table 2). DBS DNA was extracted and eluted in 50 μl of water, and 20 μl of the eluted DNA was used in the duplex assay. (A) Real-time HIV-1 amplification profiles. The HIV-1 copy numbers per ml of DBS (50 μl of whole blood per circle) are 0 (open circles), 494 (squares), 1,482 (solid triangles), 4,444 (crosses), 13,333 (open triangles), and 40,000 (solid circles). The threshold (cutoff) is 0.354. (B) Real-time RNase P amplification profiles to determine the human chromosomal DNA background of the same dried blood spots as in (A). The threshold is 0.034. (C) HIV-1 content quantification in DBS panel based on threshold cycles (C_Ts) or normalized threshold cycles (dC_Ts). The C_Ts (HIV LTR [diamonds] and RNase P [squares]) were plotted against their respective proviral copy numbers. Each datum was derived from the average of four determinations. Vertical bar: one standard deviation of C_T or dC_T. The dC_T (triangle) is the difference between the HIV C_T and its corresponding RNase P C_T.