Accurate quantification of episomal HIV-1 two-long terminal repeat circles by use of optimized DNA isolation and droplet digital PCR

J Clin Microbiol. 2015 Feb;53(2):699-701. doi: 10.1128/JCM.03087-14. Epub 2014 Dec 10.

Abstract

Episomal HIV-1 two-long terminal repeat (2-LTR) circles are considered markers for ongoing viral replication. Two sample processing procedures were compared to accurately quantify 2-LTR in patients by using droplet digital PCR (ddPCR). Here, we show that plasmid isolation with a spiked non-HIV plasmid for normalization enables more accurate 2-LTR quantification than genomic DNA isolation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA, Viral / analysis
  • DNA, Viral / isolation & purification*
  • HIV-1 / genetics*
  • Humans
  • Plasmids / analysis
  • Plasmids / isolation & purification*
  • Polymerase Chain Reaction / methods*
  • Terminal Repeat Sequences*

Substances

  • DNA, Viral