Population Genetic Analysis of the Protease Locus of Human Immunodeficiency Virus Type 1 Quasispecies Undergoing Drug Selection, Using a Denaturing Gradient-Heteroduplex Tracking Assay
- Laurence Doukhan and
- Eric Delwart *
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Fig. 1.(A) Ethidium bromide-stained DGGE gel of protease gene PCR products showing the different electrophoretic mobilities of DNA homoduplexes and heteroduplexes. Ho, position of DNA homoduplexes; He, position of DNA heteroduplexes. The numbers of mismatches in the DNA heteroduplexes are indicated above the gel. Changes in PCR products relative to that of pNL4–3 and the pairs of PCR fragments reannealed to form DNA heteroduplexes are indicated below the gel. (B) DG-HTA of plasma quasispecies collected over 7 to 14 days in three untreated subjects showing the resolved protease sequence variants and correct (i.e., reproducible) population sampling in independent nPCRs. Range of plasma viral RNA per milliliter: A lanes, 1.5 × 105to 3.5 × 105; B lanes, 2.5 × 105 to 3 × 104; C lanes, 2 × 105 to 8 × 105.
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Fig. 2.Longitudinally collected plasma viral quasispecies from P404 before and after initiation of ritonavir treatment at day 0, showing DG-HTA patterns together with Shannon entropy and the percent variant values. Viral loads were determined by branched DNA.
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Fig. 3.Longitudinally collected plasma viral quasispecies from P105 before and after initiation of ritonavir treatment at day 0, showing DG-HTA patterns together with Shannon entropy and the percent variant values. Viral loads were determined by branched DNA.
- Copyright © 2001 American Society for Microbiology
