Nucleic Acids Research, Vol 25, Issue 15 2979-2984, Copyright © 1997 by Oxford University Press
ML Collins, B Irvine, D Tyner, E Fine, C Zayati, C Chang, T Horn, D Ahle, J Detmer, LP Shen, J Kolberg, S Bushnell, MS Urdea and DD Ho
The branched DNA hybridization assay has been improved by the inclusion of
the novel nucleotides, isoC and isoG, in the amplification sequences to
prevent non-specific hybridization. The novel isoC, isoG-containing
amplification sequences have no detectable interaction with any natural DNA
sequence. The control of non-specific hybridization in turn permits
increased signal amplification. Addition of a 14 site preamplifier was
found to increase the signal/noise ratio 8-fold. A set of 74
oligonucleotide probes was designed to the consensus HIV POL sequence. The
detection limit of this new HIV branched DNA amplifier assay was
approximately 50 molecules/ml. The assay was used to measure viral load in
87 plasma samples of HIV- infected patients on triple drug therapy whose
RNA titers were <500 molecules/ml. In all 11 patients viral load
eventually declined to below the detection limit with the new assay.
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A branched DNA signal amplification assay for quantification of nucleic acid targets below 100 molecules/ml
Chiron Diagnostics, 4560 Horton Street, Emeryville, CA 94608, USA.
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