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Use of a high resolution melting (HRM) assay to compare gag, pol, and env diversity in adults with different stages of HIV infection.

  • Author(s): Cousins, Matthew M;
  • Laeyendecker, Oliver;
  • Beauchamp, Geetha;
  • Brookmeyer, Ronald;
  • Towler, William I;
  • Hudelson, Sarah E;
  • Khaki, Leila;
  • Koblin, Beryl;
  • Chesney, Margaret;
  • Moore, Richard D;
  • Kelen, Gabor D;
  • Coates, Thomas;
  • Celum, Connie;
  • Buchbinder, Susan P;
  • Seage, George R;
  • Quinn, Thomas C;
  • Donnell, Deborah;
  • Eshleman, Susan H
  • et al.
Abstract

Background

Cross-sectional assessment of HIV incidence relies on laboratory methods to discriminate between recent and non-recent HIV infection. Because HIV diversifies over time in infected individuals, HIV diversity may serve as a biomarker for assessing HIV incidence. We used a high resolution melting (HRM) diversity assay to compare HIV diversity in adults with different stages of HIV infection. This assay provides a single numeric HRM score that reflects the level of genetic diversity of HIV in a sample from an infected individual.

Methods

HIV diversity was measured in 203 adults: 20 with acute HIV infection (RNA positive, antibody negative), 116 with recent HIV infection (tested a median of 189 days after a previous negative HIV test, range 14-540 days), and 67 with non-recent HIV infection (HIV infected >2 years). HRM scores were generated for two regions in gag, one region in pol, and three regions in env.

Results

Median HRM scores were higher in non-recent infection than in recent infection for all six regions tested. In multivariate models, higher HRM scores in three of the six regions were independently associated with non-recent HIV infection.

Conclusions

The HRM diversity assay provides a simple, scalable method for measuring HIV diversity. HRM scores, which reflect the genetic diversity in a viral population, may be useful biomarkers for evaluation of HIV incidence, particularly if multiple regions of the HIV genome are examined.

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