Scheurer Michael E, Dillon Laura M, Chen Zhuo, Follen Michele, Adler-Storthz Karen
The University of Texas M. D. Anderson Cancer Center, Department of Epidemiology, Unit 1340, PO Box 301439, Houston, TX 77230-1439, USA.
Infect Agent Cancer. 2007 Apr 2;2:8. doi: 10.1186/1750-9378-2-8.
Few reports of the utilization of an accurate, cost-effective means for measuring HPV oncogene transcripts have been published. Several papers have reported the use of relative quantitation or more expensive Taqman methods. Here, we report a method of absolute quantitative real-time PCR utilizing SYBR-green fluorescence for the measurement of HPV E7 expression in cervical cytobrush specimens.
The construction of a standard curve based on the serial dilution of an E7-containing plasmid was the key for being able to accurately compare measurements between cervical samples. The assay was highly reproducible with an overall coefficient of variation of 10.4%.
The use of highly reproducible and accurate SYBR-based real-time polymerase chain reaction (PCR) assays instead of performing Taqman-type assays allows low-cost, high-throughput analysis of viral mRNA expression. The development of such assays will help in refining the current screening programs for HPV-related carcinomas.
关于利用准确、经济高效的方法测量人乳头瘤病毒（HPV）致癌基因转录本的报道很少。几篇论文报道了使用相对定量或更昂贵的Taqman方法。在此，我们报告一种利用SYBR-绿荧光进行绝对定量实时PCR的方法，用于测量宫颈细胞刷标本中HPV E7的表达。
基于含E7质粒的系列稀释构建标准曲线是能够准确比较宫颈样本测量值的关键。该检测具有高度可重复性，总体变异系数为10.4%。
使用高度可重复且准确的基于SYBR的实时聚合酶链反应（PCR）检测方法，而非进行Taqman型检测，可实现对病毒mRNA表达的低成本、高通量分析。此类检测方法的开发将有助于完善当前针对HPV相关癌症的筛查方案。
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