A Sensitive Branched DNA HIV-1 Signal Amplification Viral Load Assay with Single Day Turnaround

Branched DNA (bDNA) is a signal amplification technology used in clinical and research laboratories to quantitatively detect nucleic acids. An overnight incubation is a significant drawback of highly sensitive bDNA assays. The VERSANT® HIV-1 RNA 3.0 Assay (bDNA) (“Versant Assay”) currently used in clinical laboratories was modified to allow shorter target incubation, enabling the viral load assay to be run in a single day. To dramatically reduce the target incubation from 16–18 h to 2.5 h, composition of only the “Lysis Diluent” solution was modified. Nucleic acid probes in the assay were unchanged. Performance of the modified assay (assay in development; not commercially available) was evaluated and compared to the Versant Assay. Dilution series replicates (>950 results) were used to demonstrate that analytical sensitivity, linearity, accuracy, and precision for the shorter modified assay are comparable to the Versant Assay. HIV RNA-positive clinical specimens (n = 135) showed no significant difference in quantification between the modified assay and the Versant Assay. Equivalent relative quantification of samples of eight genotypes was demonstrated for the two assays. Elevated levels of several potentially interfering endogenous substances had no effect on quantification or specificity of the modified assay. The modified assay with drastically improved turnaround time demonstrates the viability of signal-amplifying technology, such as bDNA, as an alternative to the PCR-based assays dominating viral load monitoring in clinical laboratories. Highly sensitive bDNA assays with a single day turnaround may be ideal for laboratories with especially stringent cost, contamination, or reliability requirements

Bow shock and upstream phenomena at Mars

Zakharov, 1992). Previous investigations of planetary bow shocks have established that their position, shape and jump conditions are functions of the upstream flow parameters and the nature of the solar wind – planet interaction (Spreiter and Stahara, 1980; Slavin et al., 1983; Russell, 1985). At Mars, however, the exact nature of the solar wind interaction was elusive due to the lack of low altitude plasma and magnetic field measurements (e.g., Axford, 1991). In fact our knowledge of the nature of the interaction of Mars with the solar wind was incomplete until the arrival of MGS and the acquisition of close-in magnetic field data (Acuña et al., 1998). As detailed by a series of review papers in this monograph, the Mars Global Surveyor (MGS) mission has now shown that the Mars environment is very complex with strong, highly structured crustal magnetic remnants in the southern hemisphere, while the northern hemisphere experiences the direct impingement of solar wind plasma. This review paper first presents a survey of the observations on the Martian bow shock and the upstream phenomena in the light of results from all the missions to date. It also discusses the kinetic properties of the Martian bow shock compared to the predictions of simulations studies. Then i