Human herpesvirus 8 load in matched serum and plasma samples of patients with AIDS-associated Kaposi's sarcoma.
Human herpesvirus 8 (HHV-8) (or Kaposi's sarcoma [KS]-associated herpesvirus) is associated with all forms of KS. HHV-8 DNA load in peripheral blood mononuclear cells (PBMCs) of KS patients has been shown to correlate with the clinical stage of the disease. Studies have been done to assess the HHV-8 viral load in different sample types from KS patients and its clinical relevance. This paper describes the design and evaluation of a quantitative real-time (TaqMan) PCR assay for routine diagnosis of HHV-8 infection. The linear dynamic range was 5 to 5 x 10(6) copies of HHV-8 DNA (r(2) > 0.99). The assay is very sensitive, specific, and easily reproducible (less than 2% variability) and can be used for different clinical samples, such as serum, plasma, and PBMCs. The question of which clinical sample, serum or plasma, is preferable for HHV8 DNA testing was addressed, using this newly developed real-time PCR assay. From 85 patients with diagnosed AIDS-KS, matched plasma and serum samples were collected. Of the 85 patients tested, 35 were positive for HHV-8 DNA in both plasma and serum (41%), 8 were positive in serum but not plasma, and 7 had detectable HHV-8 DNA only in plasma. The HHV-8 load was similar in both plasma and serum, and no significant difference was found. However, more inhibition was seen in the plasma samples with the use of a system quality control, seal herpesvirus type 1. Therefore, our results suggest that serum is the preferred material for HHV-8 load testing, since there is less possible hindrance in the amplification than with plasma.
Polstra
Abeltje M
AM
Department of Human Retrovirology University of Amsterdam, The Netherlands. a.m.polstra@amc.uva.nl
Van Den Burg
Remco
R
Goudsmit
Jaap
J
Cornelissen
Marion
M
D017088:AIDS-Related Opportunistic Infections / Q000097:blood* / Q000382:microbiology; D001483:Base Sequence; D017931:DNA Primers; D004279:DNA, Viral / Q000097:blood* / Q000302:isolation & purification; D015497:HIV-1; D019288:Herpesvirus 8, Human / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods / Q000592:standards; D011786:Quality Control; D012367:RNA, Viral / Q000097:blood* / Q000302:isolation & purification; D012514:Sarcoma, Kaposi / Q000097:blood*; D019562:Viral Load / Q000592:standards
D017931:DNA Primers; D004279:DNA, Viral; D012367:RNA, Viral