Details

Autor(en) / Beteiligte

• Collins, M L
• Irvine, B
• Tyner, D
• Fine, E
• Zayati, C
• Chang, C
• Horn, T
• Ahle, D
• Detmer, J
• Shen, L P
• Kolberg, J
• Bushnell, S
• Urdea, M S
• Ho, D D

Titel

A branched DNA signal amplification assay for quantification of nucleic acid targets below 100 molecules/ml

Ist Teil von

• Nucleic acids research, 1997-08, Vol.25 (15), p.2979-2984

Ort / Verlag

England

Erscheinungsjahr

1997

Quelle

MEDLINE

Beschreibungen/Notizen

• 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.