Direct and quantitative detection of HIV-1 RNA in human plasma with a branched DNA signal amplification assay

High-Throughput Detection of West Nile Virus RNA

“…Lanciotti et al (13) recently reported a sensitive NASBA assay for WNV detection. The signal enhancement of bDNA is achieved by hybridization of viral RNA with bDNA probes (5,25). A recent comparison of the NASBA and bDNA techniques for analysis of human immunodeficiency virus RNA showed that although the bDNA test exhibited a higher dynamic range and was more robust with low copy numbers, the interassay precision of the NASBA method is better (2).…”

Quantitative, Competitive PCR Assay for HIV-1 Using a Microplate-Based Detection System

“…The remaining two plasma samples had viral loads that were below the limit of quantification by bDNA (<10 000 copies/mL), which corresponded to the QC-PCR determination of 700 and 3000 copies/mL. These results indicate that this QC-PCR technique is capable of quantifying HIV-1 virus from plasma at values comparable to those obtained from an HIV-1 viral load assay that has been shown to be reproducibly accurate (29) and that correlates well with the QC-PCR assay (2).…”

“…However, there is a growing interest in quantifying HIV-1 RNA and proviral DNA in tissues and body fluids other than peripheral blood (1,13,20,21,30). Commercially available HIV-1 viral load assays that use signal amplification (29) or the polymerase chain reaction (PCR) (17,18) were developed and are recommended for quantifying viral RNA exclusively from plasma. Here we report the development of a quantitative competitive (QC)-PCR (22)(23)(24) and a microplate detection system that uses digoxigenin (Dig)-labeled probes and an anti-Dig antibody conjugated to a bioluminescent protein (aequorin; References 5, 9 and 25-28) or peroxidase that is easy, relatively inexpensive and quantifiable to 100 copies of HIV-1 DNA or RNA per extracted sample.…”

Avances recientes en HIV/SIDA: Patogénesis, historia natural y carga viral.

“…Al agregarse un substrato quemiluminicente al plato, la reacción entre el substrato y la fosfatasa alcalina genera una cantidad de luz directamente proporcional a la cantidad de HIV-RNA presente. Se compara esta cantidad con la cantidad generada de una reacción corrida en paralelo con cantidades conocidas de RNA control y se puede llegar de esta menera a determinar la cantidad inicial de HIV-RNA presente en la muestra de plasma del paciente (18).…”