Abstract
An extremely sensitive and convenient microtitre plate solution hybridisation assay for the detection of HIV-1 PCR products was developed. The PCR product is labelled by direct incorporation of digoxigenin-dUTP and after denaturation is captured by a microtitre plate coated with a streptavidin-linked biotinylated probe. The PCR/probe hybrids are reacted with an alkaline phosphatase conjugated anti-digoxigenin antibody and detected using an alkaline phosphatase enzyme amplification system. The use of uracil-N-glycosylase and dUTP instead of dTTP in the PCR is used to effectively control carry-over from previous PCR products. The assay can detect single HIV-1 DNA molecules in a background DNA of 0.75μg.
| Original language | English |
|---|---|
| Pages (from-to) | 67-76 |
| Number of pages | 10 |
| Journal | Journal of Virological Methods |
| Volume | 44 |
| Issue number | 1 |
| DOIs | |
| Publication status | Published - 1 Sept 1993 |
| Externally published | Yes |
UN SDGs
This output contributes to the following UN Sustainable Development Goals (SDGs)
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SDG 3 Good Health and Well-being
Keywords
- Digoxigenin
- HIV-1
- Hybridisation
- PCR
- UNG
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