Multicenter Comparison of PCR Assays for Detection of Human Herpesvirus 8 DNA in Semen

Saliva samples were collected from both mothers and children (for infants, saliva was collected by use of a pipette). Virus from 180 ml supernatant was pelleted, and the pellet was resuspended in 1,000 μl PBS, of which 50 μl was injected per implant. [16], with similar performance characteristics. Thus, the final study group consisted of 2750 participants (81% of the ZAPP cohort). The publication costs of this article were defrayed in part by page charge payment. This is supported by earlier reports of high incidence rates of classic KS in this region [20].

Other data—such as the race or ethnicity of the recipient, age and sex of the donor, type of immunosuppressive therapy, clinical outcome of transplant recipient, and the number of blood transfusions each recipient received—were not available for a sufficient number of the subjects to permit analyses. As part of our continuing cohort study of the epidemiology of HHV-8 infection among Zambian mother-infant pairs, the major goal of the present study was to examine whether transmission of HHV-8 to infants can occur via breast milk. The low percentage of transplants is a consequence of a scarcity of donors in central and southern Italy. Details of the study design, rationale, and recruitment process for this study have been previously described in detail (22). Further HHV-8 transmission studies in the HIV-seronegative population demonstrate additional data in the transmission, development, and persistence of HHV-8-associated malignancy. A real-time (TaqMan) quantitative PCR on a conserved region of ORF26 was then performed, as previously described (22).

To detect HHV-8, we used two sets of nested primers specific for two nonoverlapping genomic regions. The supernatant fractions were all negative for HHV-8 with both primer sets in unnested and nested PCRs. This pattern suggests that the primary mode of KSHV transmission in this region is horizontal and nonsexual during childhood and that there is ongoing transmission throughout childhood. IFA using rSFV K8.1-infected BHK21 cells (K8.1SFV IFA) was as sensitive as lytic PEL-based IFA, and the correlation coefficient of end-point titers and concordance between these two assays were >0.92 and 97% (κ = 0.93), respectively. A). [19] with modifications.

Blood was collected and subjects were interviewed at enrollment and about every 12 months thereafter. Since it is well known that gonococcal uretrithis is efficiently acquired through insertive oral sex among homosexual and heterosexual males, the finding that prevalent HHV-8 infection tended to be associated with gonorrhea in both groups may indicate that this STI could be a biological marker of exposure to saliva during sex in both groups. Both the detection of HHV-8 DNA in the saliva of infected children [27] and the association between children’s infection status and their mother’s higher saliva HHV-8 loads [28] are consistent with the occurrence of HHV-8 transmission, via saliva, within families. For serologic testing for antibodies to HHV-8 and other viruses, serum samples were stored at -20°C in the laboratories of Tor Vergata University, Rome, Italy. Three kidney transplant recipients who developed KS 17 months to 7 years after transplantation were examined. As Director of this program, based at the Fred Hutchinson Cancer Research Center, he established a translational clinical research site in Kampala.

The association of KS with groups that acquired human immunodeficiency virus type 1 (HIV-1) sexually led investigators to speculate that there was a sexually transmitted agent related to KS. Data from their observational cohort study of 1092 transfusion recipients of known HHV-8–seropositive or HHV-8–seronegative blood in 1 hospital in Uganda, from late 2000 to mid-2001, showed that 17% of recipients of HHV-8–seropositive blood stored for ≤4 days (short-stored) died 1 week to 6 months posttransfusion compared with 10% of recipients of seropositive blood stored >4 days (long-stored) and 8% of recipients of seronegative blood. Maternal HIV-1 and HHV-8 infection status were not independently associated with risk of HHV-8 seroconversion in the child. The presence of anti-HHV-8 antibodies in serum samples was investigated by an immunfluorescence assay. His immunology work described the first comprehensively-validated serologic assay for detection of antibodies to HHV-8, the original description of neutralizing antibodies to HHV-8 and their role in development of Kaposi Sarcoma (KS), and the characterization of gamma-delta T-cell activity in persons with chronic HHV-infection. Of these APC, only B cells support complete, lytic HHV-8 infection.