No separated sample had a viral load > 200 copies/mL. Only two whole-blood

samples had a viral load of < 40 copies/mL compared with 19 of 21 separated samples (90%). All separated samples had an HIV-1 viral load of 54 copies/mL or less, i.e. nil had a significant viraemia (Fig. 1). The range of results for whole-blood samples was from ‘not detected’ to 3080 copies/mL; the mean was 629 copies/mL and the median 279 copies/mL. Further research in this important area is needed. HIV-1 RNA results that are above the cut-off in patients on treatment have much greater implications than a slightly inaccurate result in a patient off treatment. There is currently no evidence in the MK-2206 mouse literature which relates to the reproducibility of HIV RNA assays at low copy number relating to different periods of time pre-centrifugation in patients on ART. Therefore, until these data become available using current assays, including Roche TaqMan v2.0, we

suggest that plasma separation should occur at under 24 hours, ideally at under 8 hours. Close attention needs to be paid to the timing of plasma separation in patients on ART who are pregnant and enrolled in clinical trials. “
“The aim of this study was to develop a system for rapid and accurate real-time quantitative PCR (qPCR) identification and quantification of Botrytis cinerea, one of the major pathogens present on grapes. The intergenic spacer (IGS) region of the nuclear ribosomal DNA was used to specifically detect and quantify Akt inhibitor B. cinerea. A

standard curve was established to quantify this fungus. The qPCR reaction was based on the simultaneous detection of Farnesyltransferase a specific IGS sequence and also contained an internal amplification control to compensate for variations in DNA extraction and the various compounds from grapes that inhibit PCR. In these conditions, the assay had high efficiency (97%), and the limit of detection was estimated to be 6.3 pg DNA (corresponding to 540 spores). Our method was applied to assess the effects of various treatment strategies against Botrytis in the vineyard. Our qPCR assay proved to be rapid, selective and sensitive and may be used to monitor Botrytis infection in vineyards. Many fungal and bacterial organisms, of which Botrytis cinerea is the most important, can infect grapes and cause a ‘bunch rot’ (Keller et al., 2003). The disease caused by B. cinerea, also known as ‘grey mould’, is arguably the most significant disease problem confronting the wine industry worldwide. The presence of grey mould on grapes is undesirable, as it lowers the quality of wines. Depending on the vintage, fungal infection rates can reach 15–25% of grapes, and wines prepared from infected grapes usually exhibit organoleptic defects, such as colour oxidation or the appearance of typical aromatic notes (‘moldy’, ‘rotten’), which are not appreciated by consumers (Cilindre et al., 2007).