The purpose of the Laboratory Core is to provide virology laboratory support and services for the three Research Projects. Specifically, these services include screening to determine eligibility requirements, supervising the collection of specimens for monitoring virological markers of disease progression and regression of CMV, HSV and HIV. These studies will be conducted through the Laboratory Core at the University of Washington, the University of Rochester and the University of Nairobi. In addition, should the need arise over the course of the Program Project. Viral DNA detection procedures for studies of opportunistic infections in HIV-infected women due to the hepatitis C virus herpes virus type 6 (HHV-6) and type 8 (HHV-8/HKSV) are available in the Virology Division at the University of Washington. A diverse virological laboratory base is important for elucidating the pathogenesis of HIV infection in women. The specific goals of the Clinical Retrovirology Laboratory Core are as follows: 1. To coordinate the clinical virology activities of the University of Washington and affiliated institutions for the development and support of complementary research projects in HIV-1 shedding and diversity (Project I: Drs. Frenkel, Coombs and Mullins), CMV as a co-factor for HIV-1 shedding (Project III. Drs. Hitti, Cohn and Cohen). 2. To maintain and develop the facilities, equipment and staffing needed for an infrastructure to accommodate these complementary research projects. Virological procedures to accomplish the above purpose and goals are certified (NIAID-sponsored Virology Quality Assurance Program; CDC; CAP) or peer reviewed and include: i) HIV-1/2 antibody assays include EIA to detect antibodies to viral antigens and recombinant antigens; immunoblotting to detect antibodies to specific HIV proteins such as p24, gp41 and gp120/160; HSV-1/-2 antibody detection (screening). ii) Quantitative determinations of HIV-1 RNA in body tissues, oral fluid, genital secretions and blood (Projects I, II and III). iii) Determination of HIV total DNA by polymerase chain reaction (PCR) amplification using a novel quantitative-competitive PCR-EIA (QC-PCR- EIA) and by a Taq polymerase-based real-time PCR assay (Project I, II, III). iv) Quantification of HIV-1 unintegrated episomal 2-LTR DNA by in situ PCR and in situ hybridization (ISH) (Project II). Vi) Detection and quantification of CMV, HCV, and HSV DNA by TaqMan PCR (Project II).
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