Nucleic acid assay system for tier II laboratories and moderately complex clinics to detect HIV in low-resource settings.
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.
Tang
Wen
W
BioHelix Corporation, Beverly, Massachusetts, USA.
Chow
Wing Huen A
WH
Li
Ying
Y
Kong
Huimin
H
Tang
Yi-Wei
YW
Lemieux
Bertrand
B
D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification; D006678:HIV / Q000235:genetics / Q000302:isolation & purification*; D015658:HIV Infections / Q000175:diagnosis* / Q000821:virology*; D006801:Humans; D007118:Immunoassay / Q000379:methods; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification; D012680:Sensitivity and Specificity
D004279:DNA, Viral; D012367:RNA, Viral