The molecular diagnostics laboratory processed approximately 900 unique clinical samples in 2007, accounting for over 3000 tests. Approximately 300 additional tests were performed to support a variety of translational research projects, for NCI/NIH PIs. The majority of the laboratory's tests are conventional PCR assays, including both DNA and RT-PCR assays. The assays performed by the molecular diagnostics laboratory include assays for B and T-cell clonality, assays for lymphoma related translocations, assays for cancer associated viruses (EBV, HTLV, HHV-8), and RT-PCR assays for translocations associated with pediatric sarcomas. Quantitative PCR is performed for HTLV 1/2 to follow viral load for the Metabolism Branch's ATL clinical trials. Assays are performed on materials derived from a variety of sources. Paraffin fixed tissue specimens account for about 60% of the samples and blood, 30%. The remaining 10% of samples are derived from fresh or frozen tissue, cerebrospinal fluid, and other body fluids. A high percentage of the paraffin section studies are complex and labor intensive, requiring macro or microdissection of specific components of lesional tissues, and are performed under the guidance of a fellow or staff pathologist. The molecular laboratory is particularly expert in extracting nucleic acids from small fragments of archival paraffin-embedded tissues, and our expertise in this area has been widely sought out in a number of laboratories within and outside of NIH. The laboratory maintains 3 Perkin-Elmer 9700 PCR platforms, one ABI 7900 real-time PCR machine, and one ABI 3130 capillary electrophoresis unit. As part of the recent funding initiative to expand molecular diagnostics testing in the LP, the laboratory just purchased a Pyromark 2400 pyrosequencer which will be employed initially for K-RAS mutation analysis, and for MGMT promoter methylation analysis. The molecular diagnostics laboratory supports translational research of both NCI and NIH researchers. Among the NCI and NIH tenured investigators and clinicians that have utilized the laboratory's resources and data to support their publications are: Dr. Elaine Jaffe (NCI) Dr. Maria Tsokos (NCI) Dr. Andrea Abati (NCI) Dr. Robert Kreitman (NCI) Dr. Wyndham Wilson (NCI) Dr. John Janik (NCI) Dr. John Morris (NCI) Dr. Steven Holland (NIAID) Dr, Richard Little (NCI) Dr. Dean Metcalfe (NIAID) Dr. Amy Klion Dr. John Morris
