Development of a quantitative competitive polymerase chain reaction for human herpesvirus 8

J Virol Methods. 1997 Feb;64(1):19-26. doi: 10.1016/s0166-0934(96)02139-8.

Abstract

A quantitative competitive polymerase chain reaction (PCR) assay for human herpesvirus 8 (HHV8) was developed. The assay uses a competitive internal control sequence which differs from the wild type sequence by the presence of an EcoRI restriction endonuclease site. The quantitative method was validated by co-amplifying known amounts of control sequence DNA and wild type DNA, and shown to accurately quantify HHV8 within the range of 10(1)-10(6) genome copies (R = 0.998). The assay was used to quantify HHV8 in sequential blood samples of a renal transplant patient diagnosed with Kaposi's sarcoma (KS). A peak viral load of 28320 genomes/ml blood was present at diagnosis of KS, which reduced to 13680 genomes after 2 days and was undetectable 4 months later. Control of HHV8 replication in this patient parallelled the cessation of immunosuppressive therapy.

MeSH terms

  • Calibration
  • DNA, Viral / analysis*
  • Follow-Up Studies
  • Herpesvirus 8, Human / genetics
  • Herpesvirus 8, Human / isolation & purification*
  • Humans
  • Kidney Transplantation
  • Plasmids
  • Polymerase Chain Reaction / methods*
  • Sarcoma, Kaposi / blood
  • Sarcoma, Kaposi / virology*

Substances

  • DNA, Viral