Nucleic Acid Assay System for Tier II Laboratories and Moderately Complex Clinics to Detect HIV in Low-Resource Settings

J Infect Dis. 2010 Apr 15;201 Suppl 1(Suppl 1):S46-51. doi: 10.1086/650388.


There is a clear need for an instrument-free molecular diagnostic system for detecting human immunodeficiency virus type 1 (HIV-1) RNA or DNA that can be used in developing countries. Such a test could be used for early diagnosis of HIV-1 infection during infancy and could serve as a surrogate end point for vaccine trials. We developed the IsoAmp HIV-1 assay (BioHelix Corporation), which targets the HIV-1 gag gene with use of isothermal reverse-transcription helicase-dependent amplification chemistry. The IsoAmp HIV assay uses a disposable amplicon containment device with an embedded vertical-flow DNA detection strip to detect the presence of HIV-1 amplicons. The vertical-flow DNA detection strip has a control line to validate the performance of the device and a test line to detect the analyte. The analyte is detected by a sandwich immunoassay for reporter moieties on a capture probe and a detection probe. The control line consists of the detection probe reporter moiety conjugated to the vertical-flow DNA detection strip. The preliminary limit of detection of the IsoAmp HIV assay was evaluated by testing serial dilutions of HIV-1 armored RNA (Assuragen). We found that 21 (75%) of 28 assays yielded positive results when 50 copies of HIV-1 armored RNA were input into the IsoAmp HIV reaction.

Publication types

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

MeSH terms

  • DNA, Viral / genetics
  • DNA, Viral / isolation & purification
  • HIV / genetics
  • HIV / isolation & purification*
  • HIV Infections / diagnosis*
  • HIV Infections / virology*
  • Humans
  • Immunoassay / methods
  • Molecular Diagnostic Techniques / methods*
  • Nucleic Acid Amplification Techniques / methods*
  • RNA, Viral / genetics
  • RNA, Viral / isolation & purification
  • Sensitivity and Specificity


  • DNA, Viral
  • RNA, Viral