RESUMEN
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.
Asunto(s)
ADN/química , Hibridación de Ácido Nucleico/métodos , Adenosina , Fármacos Anti-VIH/uso terapéutico , Citidina/química , ADN/metabolismo , ADN Viral/análisis , Quimioterapia Combinada , Guanosina/química , VIH , Infecciones por VIH/tratamiento farmacológico , Infecciones por VIH/virología , Humanos , Isoquinolinas/uso terapéutico , Lamivudine/uso terapéutico , Nelfinavir , ARN Viral/análisis , Ácidos Sulfónicos/uso terapéutico , Zidovudina/uso terapéuticoRESUMEN
Analogues of bradykinin were synthesized containing single substitutions by N alpha-methyl amino acids in the 1, 4, 5, 5, and 9 positions. [MeArg]Bradykinin possessed 60% of the muscle-contracting activity of the parent compound in a guinea pig ileum assay. The other analogues were very weak agonists (less than 2%) and, disappointingly, failed to show blocking activity except at very high doses.