UNIVERSITY OF PITTSBURGH THE
The purpose of the administrative supplement is to request funds to purchase a Real time Taqman instrument which will be used in studies related to the parent grant. Currently, our laboratory does not have a real time PCR machine and we are using ABI 7000, which is in our collaborator's laboratory. This instrument does not have the capabilities to read 384 well plates and the large number of samples l. Additionally, this equipment is heavily utilized by several other investigators in our Department. It is not unusual that students and staff from our laboratory have to wait in order to have their samples analyzed. In addition, this Taqman machine is very old and cannot be used for multiprobe detection systems and has been requiring frequent expensive repairs. The new opportunities provided by the Administrative supplement will increase the value of the parent grant by allowing improved high sensitive measurements of viral load in semen and anatomical parts of the male genital organ. This requested instrument will be especially useful in measuring viral load after therapy in semen and blood, also in anatomical parts of the male genital organ where viral load is expected to be extremely low. As an extension we will investigate the presence of CMV in semen. Presence of CMV in semen has been implicated to increase inflammation in genital organ and thereby increase HIV shedding in semen. Availability of a newer the ABI-Prism 7900HT with 384 plate capacity will allow simultaneous measurement of CMV and HIV-1 in semen using multiplexing technology. In addition, ABI 7900HT utilizes smaller sample volume with high sensitivity, which is a key factor in dealing with semen which is often obtained in small volume. This machine will also be used for future HIV transmission studies.