BACKGROUND Human herpesvirus 8 (HHV-8) virological diagnosis and monitoring relies mainly on real-time PCR. OBJECTIVES To evaluate two real-time PCR commercial kit (HHV-8 Premix R-geneTM and Clonit® HHV-8) and compare with in-house real-time PCR. STUDY DESIGN Twelve samples (3 undetectable and 9 detectable with viral load ranging from 101 to 105 per reaction) were tested for HHV-8 detection and quantification with the 3 methods. Methods comparison was supported with regression curve and diagram presenting difference or ratio between commercial and in-house PCR results and plotted against the in-house PCR results. Statistical analyses, specifically Student tests and Spearman correlation, were performed. RESULTS In both cases, qualitative results obtained with commercial kit and in-house PCR were identical and HHV-8 quantitation results were significantly correlated (Clonit®, Rs = 1, p < 0.001 and R-geneTM Rs = 0.98, p < 0.001). However, Clonit® results were significantly higher compared to the in-house results with an overestimation in median [IQR] of 1.16 log10 copies/106 cells [1.12-1.18] whereas R-GeneTM results were not significantly higher, and an overestimation in median of 0.46 log10 copies/106 cells [0.37-0.52]. Otherwise, repeatability and reproducibility tests of undetectable sample failed with Clonit® technique contrary to the R-GeneTM. CONCLUSIONS HHV-8 R-geneTM assay seems to be the most suitable since it showed consistent qualitative results with in-house HHV-8 PCR, a good quantitative correlation, an overestimation not significantly different and inferior to 0.50 log10 copies/106 cells and a good repeatability.

中文翻译：

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人疱疹病毒8感染的分子诊断方法比较。

背景技术人类疱疹病毒8（HHV-8）病毒学诊断和监测主要依赖于实时PCR。目的评估两种实时PCR商业试剂盒（HHV-8 Premix R-geneTM和Clonit®HHV-8），并与内部实时PCR进行比较。研究设计测试了12种样品（3种未检出样品和9种可检出样品，每个反应的病毒载量范围为101至105），用3种方法测试HHV-8的检测和定量。方法比较得到回归曲线和图表的支持，该曲线和图表显示了商业PCR和内部PCR结果之间的差异或比率，并针对内部PCR结果进行了绘制。进行统计分析，特别是学生测试和Spearman相关性。结果在两种情况下，使用商业试剂盒和内部PCR获得的定性结果相同，并且HHV-8定量结果显着相关（Rs = 1，p <0.001和R-geneTM Rs = 0.98，p <0.001）。但是，与内部结果相比，Clonit®结果明显更高，其中中位数[IQR]被高估了1.16 log10个拷贝/ 106个细胞[1.12-1.18]，而R-GeneTM结果并未显着更高，并且中值被高估了0.46 log10拷贝/ 106个单元[0.37-0.52]。否则，与R-GeneTM相反，采用Clonit®技术无法对无法检测到的样品进行重复性和再现性测试。结论HHV-8 R-geneTM检测似乎是最合适的，因为它与内部HHV-8 PCR表现出一致的定性结果，具有良好的定量相关性，