Lee TH, Chrebtow V, Busch MP; Conference on Retroviruses and Opportunistic Infections.
Program Abstr 6th Conf Retrovir Oppor Infect Conf Retrovir Oppor Infect 6th 1999 Chic Ill. 1999 Jan 31-Feb 4; 6th: 106 (abstract no. 203).
Blood Centers of the Pacific, San Francisco CA.
Background: HHV-8 has been shown to be associated with development of Kaposi's Sarcoma (KS) in HIV+ individuals; however, the relationship between the HHV-8 viral load in blood and transmission or development of KS is unclear. We developed a quantitative PCR assay to characterize viral load of HHV-8 in white blood cells (WBC). Method: 0.5mL of fresh and frozen/thawed whole blood samples were processed into WBC pellets by saponin lysis buffer, followed by digestion, amplification and detection by 32P-labeled probe. HHV-8 sequence-specific primers (KS or GBC) were used in amplification. A serial dilution of BCBL-1 lysate was included in each run as an external standard curve. Internal control (IC) DNA, which includes KS or GBC primer sites but is detected by different unique probes, was spiked into amplification reaction mixture for control of amplification inhibition. Results: The sensitivity of the PCR assays are approximately 4-20 HHV-8 genome equivalents (ge) per mL blood and the dynamic range approximately 4 logs, based on amplification of dilutions of BCBL-1 lysate (estimated 50 copies/cell). One of 100 normal blood donor samples, one of 60 female AIDS patients, and 13 of 100 male HIV-1+ persons were positive for HHV-8 by the PCR assay (Table). Results were similar for fresh and frozen blood. (Table: see text) Conclusions: We have developed a sensitive quantitative PCR assay for HHV-8. This assay should be useful for characterizing HHV-8 viral load in natural history studies and for monitoring viral load after drug therapy.
Publication Types:
Keywords:
- Acquired Immunodeficiency Syndrome
- Base Sequence
- Female
- HIV Infections
- HIV Seropositivity
- HIV-1
- Herpesvirus 8, Human
- Humans
- Male
- Polymerase Chain Reaction
- Sarcoma, Kaposi
- Sensitivity and Specificity
- Serologic Tests
- Viral Load
- genetics
- organization & administration
Other ID:
UI: 102188845
From Meeting Abstracts