Detection of HIV-1 Drug Resistance in Women Following Administration of a Single Dose of Nevirapine: Comparison of Plasma RNA to Cellular DNA by Consensus Sequencing and by Oligonucleotide Ligation Assay

J Clin Microbiol. 2010 May;48(5):1555-61. doi: 10.1128/JCM.02062-09. Epub 2010 Feb 24.

Abstract

A single dose of nevirapine (sdNVP) to prevent mother-to-child transmission of HIV-1 increases the risk of failure of subsequent NVP-containing antiretroviral therapy (ART), especially when initiated within 6 months of sdNVP administration, emphasizing the importance of understanding the decay of nevirapine-resistant mutants. Nevirapine-resistant HIV-1 genotypes (with the mutations K103N, Y181C, and/or G190A) from 21 women were evaluated 10 days and 6 weeks after sdNVP administration and at the initiation of ART. Resistance was assayed by consensus sequencing and by a more sensitive assay (oligonucleotide ligation assay [OLA]) using plasma-derived HIV-1 RNA and cell-associated HIV-1 DNA. OLA detected nevirapine resistance in more specimens than consensus sequencing did (63% versus 33%, P<0.01). When resistance was detected only by OLA (n=45), the median mutant concentration was 18%, compared to 61% when detected by both sequencing and OLA (n=51) (P<0.0001). The proportion of women whose nevirapine resistance was detected by OLA 10 days after sdNVP administration was higher when we tested their HIV-1 RNA (95%) than when we tested their HIV-1 DNA (88%), whereas at 6 weeks after sdNVP therapy, the proportion was greater with DNA (85%) than with RNA (67%) and remained higher with DNA (33%) than with RNA (11%) at the initiation of antiretroviral treatment (median, 45 weeks after sdNVP therapy). Fourteen women started NVP-ART more than 6 months after sdNVP therapy; resistance was detected by OLA in 14% of the women but only in their DNA. HIV-1 resistance to NVP following sdNVP therapy persists longer in cellular DNA than in plasma RNA, as determined by a sensitive assay using sufficient copies of virus, suggesting that DNA may be superior to RNA for detecting resistance at the initiation of ART.

Publication types

  • Comparative Study
  • Evaluation Study
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Anti-HIV Agents / therapeutic use*
  • Chemoprevention / methods
  • DNA Primers / genetics
  • DNA, Viral / blood
  • Drug Resistance, Viral*
  • Female
  • HIV Infections / drug therapy
  • HIV-1 / genetics*
  • HIV-1 / isolation & purification
  • Humans
  • Infectious Disease Transmission, Vertical / prevention & control*
  • Ligase Chain Reaction / methods
  • Microbial Sensitivity Tests / methods
  • Nevirapine / therapeutic use*
  • Oligonucleotide Probes / genetics
  • Pregnancy
  • Pregnancy Complications, Infectious / drug therapy
  • RNA, Viral / blood
  • Sensitivity and Specificity
  • Sequence Analysis, DNA / methods

Substances

  • Anti-HIV Agents
  • DNA Primers
  • DNA, Viral
  • Oligonucleotide Probes
  • RNA, Viral
  • Nevirapine