Lock M J, Griffiths P D, Emery V C
Department of Virology, Royal Free Hospital School of Medicine, University of London, UK.
J Virol Methods. 1997 Feb;64(1):19-26. doi: 10.1016/s0166-0934(96)02139-8.
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.
开发了一种用于检测人类疱疹病毒8型（HHV8）的定量竞争性聚合酶链反应（PCR）分析方法。该分析方法使用了一个竞争性内部对照序列，该序列因存在一个EcoRI限制性内切酶位点而与野生型序列不同。通过共同扩增已知量的对照序列DNA和野生型DNA对定量方法进行了验证，结果表明该方法能在10(1)-10(6)个基因组拷贝范围内准确量化HHV8（R = 0.998）。该分析方法用于对一名被诊断为卡波西肉瘤（KS）的肾移植患者的连续血样中的HHV8进行定量。在诊断为KS时，血液中病毒载量峰值为28320个基因组/毫升，2天后降至13680个基因组，4个月后检测不到。该患者体内HHV8复制的控制与免疫抑制治疗的停止情况平行。
