Evaluation of a qualitative human immunodeficiency virus-1 diagnostic assay based on nucleic acid sequence based amplification and lateral flow readout

摘要

In this study, a qualitative human immunodeficiency virus-1 (HIV-1) diagnostic assay was developed and evaluated as part of an effort to create a point-of-care diagnostic test for pediatric HIV-1. The assay is based on extraction of viral ribonucleic acid (RNA) with magnetic microbeads, isothermal amplification using nucleic acid sequence based amplification (NASBA), and detection of amplified RNA on lateral flow strips. Using mock samples consisting of plasma spiked with in vitro transcribed RNA or HIV-1C particles, more than 80% of samples with a concentration of 1,000 copies/mL or higher tested positive with the assay. Using clinical samples from pediatric patients at the Queen Elizabeth Central Hospital in Blantyre, Malawi, more than 80% of samples with a concentration of 10,000 copies/mL or higher tested positive. These results indicate that this assay is capable of detecting HIV-1 at concentrations found in pediatric samples and shows promise for use in low resource settings but requires further improvements to increase the specificity and sensitivity at low viral loads.