Project Description

USA clinical laboratory needed for HIV plasma testing: detect virus in human plasma using TEM Need to visualize HIV particles in human plasma using TEM to correlate with viral load measurements. Must detect HIV by size and morphology only. Have successfully used TEM to detect two different clones of HIV added to plasma at concentrations of 1 million particles/ml and above. Must be able to detect virus at concentrations typical of viral load numbers, which range from 10 to 1000-fold below the practical limit of TEM detection. Centrifugation alone does not work. Steric exclusion HPLC can purify retroviruses in blood plasma and achieve over a 1100-fold purification. HIV elutes in the void volume. Centrifuge void volume to a pellet, suspend in 20 uL buffer, do TEM. (Darling et al. (1977) J Chromatogr 131:383-390.)