The Virology Laboratory Core provides the HSV virus culture service, directed by Drs. Corey and Ashley, includes virus isolation and antigen detection to identify infectious HSV in genital lesions and in oral, genital and rectal secretions (Projects I, III, IV). All women delivering infants at the two participating hospitals are sampled at delivery for virus isolation provided by the Laboratory Core (Project II). The Core will also provide antiviral sensitivity testing on subsets of isolates from Projects I and III. HSV type specific Western blot service, directed by Dr. Ashley, is provided for on prenatal and delivery serum pairs to identify women seroconverting to HSV during pregnancy (Project II). This test is also provided in support of Projects I, III, and IV to determine serostatus for subject cohorting as: seronegative for HSV, HSV-1 seropositive, HSV-2 seropositive or seropositive for both HSV- 1 and HSV-2. These services will be centralized in a single location at CHMC along with the isolate and serum banks and their respective computer databases which interface with the facilities of the Project Database Manager. In addition, HSV PCR services, directed by Dr. Cone, include diagnosis of low titer viral shedding by detection of amplified HSV DNA, determination of actively replicating HSV by detection of amplified HSV RNA, quantitative RNA and quantitative DNA to provide measures of viral load at sites of HSV shedding. In selected cases, restriction enzyme and sequence analyses will be performed in conjunction with the HSV-PCR service. In addition to laboratory service work relating to HSV diagnosis, an HIV service component, directed by Drs. Corey and Coombs, is included in the Laboratory Core in support of Project III. The tests provided are all certified by the AIDS Collaborative Treatment Group and/or the College of American Pathologists and include: HIV antibody binding and neutralizing assays in cell lines and in PBMCs, p24 antigen determinations, measurement of mucosal antibodies by chemiluminescence immunoblot, HIV isolation, and quantitation, determination of fusogenic characteristics of isolates, antiviral sensitivity testing of isolates, detection and quantitation of HIV RNA by PCR and branched chain PCR and detection and quantitation of DNA, and DNA replicative intermediates by PCR. All methods described in the Virology Laboratory Core have exhaustive integral quality control measures and are used routinely in the laboratories of the faculty who will direct the respective support services for the Program Project.
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