Quantitation of cell-free and cell-associated Kaposi's sarcoma-associated herpesvirus DNA by real-time PCR.
A real-time PCR assay for quantitation of Kaposi's sarcoma-associated herpes virus (KSHV or human herpesvirus 8) DNA was evaluated. The linear dynamic range was 10 to 10(5) copies of KSHV DNA (r(2) > 0.99). The accuracy of DNA purification and quantitation was less than +/-0.4 log(10) copies for samples that contained from 10 to 10(5) copies of KSHV DNA. Cell-associated KSHV DNA was quantitated over a range of infected cell frequencies from 0. 1 to 10(-5), and cell-free KSHV DNA in plasma was quantitated over a range of 100 to 10(6) copies/ml. Real-time PCR provides a convenient method for quantitation of cell-free and cell-associated KSHV DNA in laboratory and clinical specimens.
White
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IE
Division of Infectious Diseases, Department of Medicine, University of Colorado Health Sciences Center, Denver, Colorado, USA.
Campbell
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TB
D002460:Cell Line; D004279:DNA, Viral / Q000097:blood; D019288:Herpesvirus 8, Human / Q000302:isolation & purification* / Q000502:physiology; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods; D015203:Reproducibility of Results; D012514:Sarcoma, Kaposi / Q000097:blood / Q000175:diagnosis*; D012680:Sensitivity and Specificity; D014771:Virion / Q000502:physiology; D014779:Virus Replication
D004279:DNA, Viral