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Versant Branched-DNA Assay for Reliable Detection of HIV RNA Using Alkaline Phosphatase

The Versant Branched-DNA Assay leverages alkaline phosphatase pre-amplifier and target probes to detect HIV RNA from lysed particles in a 96-well opaque format. This assay demonstrates a direct correlation between the number of RNA strands and the intensity of chemiluminescence generated, with detection thresholds as low as 450 copies of HIV RNA/mL. Performance results show variations, with high sensitivity of ~734,000 cp/mL and a reliable lower limit of detection at ~450 copies/mL, challenging comparisons with the Wave80 assay.

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Versant Branched-DNA Assay for Reliable Detection of HIV RNA Using Alkaline Phosphatase

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  1. Versant Branched-DNA assay Label probes + Alkaline phophatase Pre-amplifier probe Amplifier probes Target probe HIV RNA (from lysed particles) Capture probe Opaque 96-well • # of RNA strands is proportional to chemiluminescence intensity from • alkaline phosphatase reaction

  2. Wave80’s Versant Branched-DNA assay A ~ 734,000 cp/mL B ~ 45,000 cp/mL C ~ 4,500 copies/mL E ~ 90 copies/mL D ~ 450 copies/mL Neg. control • Reliable detection threshold ~ 450 copies HIV RNA/mL

  3. Our b-DNA assay has lower sensitivity than Wave80 A Hi B C C ~ 4,500 copies/mL Lo D E Neg. ctrl F • Plate reader either has lower sensitivity / dynamic range, or we could have • overall RNA breakdown

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