Intra- and inter-laboratory variability in human T-cell leukemia virus type-1 proviral load quantification using real-time polymerase chain reaction assays: A multi-center study
Intra- and inter-laboratory variability in human T-cell leukemia virus type-1 proviral load quantification using real-time polymerase chain reaction assays: A multi-center study
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DOI：
10.1111/j.1349-7006.2010.01720.x
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发表时间：
2010-11-01
期刊：
影响因子：
5.7
通讯作者：
Watanabe, Toshiki
中科院分区：
文献类型：
--
作者：
Kamihira, Shimeru;Yamano, Yoshihisa;Watanabe, Toshiki
人类T细胞白血病病毒1型（HTLV-1）前病毒载量（VL）是病毒致病和恶性演变的重要决定因素。虽然VL已经通过内部实时定量聚合酶链反应（qPCR）技术进行了定量，但对不同VL检测系统之间的协调性知之甚少。我们评估了实验室内和实验室间的变异性VL测量在六个实验室使用相同的DNA样本血清阳性的HTLV-1在两个步骤的方式。第一项研究通过原始内部测定法测量了60份样本，发现实验室内和实验室间的中位变异系数（CV）分别为44.9%（范围：25.4-71.8%）和59.9%（34.2-93.4%）。实验室间相关系数范围为0.760至0.875，表明每个实验室的VL测量精度良好，但实验室间回归斜率在0.399至2.206之间，表明VL测量在实验室之间存在较大差异。为了检查参考物质（RM）标准化对VL变异性的影响，我们仅使用20份样品进行了第二项研究，用RM替代了包含HTLV-1 pX区域的质粒。原始pX拷贝数的中位实验室间CV从66.9%显著降低至35.7%，而内部对照的中位CV几乎保持不变，导致VL的实验室间CV无改善。这表明，每个内部测定系统工作良好，具有良好的精密度，但单独标准化RM不足以实现协调。不仅RM，而且用于数据标准化的内部对照基因的相关选择预期对于标准化HTLV-1 VL测量是现实的。（Cancer Sci 2010; 101：2361-2367）。
Human T-cell leukemia virus type-1 (HTLV-1) proviral load (VL) is an important determinant of viral pathogenesis and malignant evolution. Although VL has been quantified by in-house real-time quantifiable polymerase chain reaction (qPCR) technology, little is known about the harmonization among different VL assay systems. We evaluated intra- and inter-laboratory variability of VL measured at six laboratories using the same DNA samples seropositive for HTLV-1 in a two-step manner. The first study measured 60 samples by original in-house assays, finding that the median intra- and inter-laboratory coefficient of variation (CV) was 44.9% (range, 25.4-71.8%) and 59.9% (34.2-93.4%), respectively. The inter-laboratory correlation coefficients ranged from 0.760 to 0.875, indicating that VL were measured with good precision in each laboratory, but inter-laboratory regression slopes differed from 0.399 to 2.206, indicating that VL were measured with a wide variation between laboratories. To examine the effect of standardization of reference materials (RM) on the VL variability, we performed a second study using only 20 samples by substituting RM for plasmid including the HTLV-1 pX region. The median inter-laboratory CV for raw pX copy number was reduced significantly from 66.9% to 35.7%, whereas the median CV for the internal control remained almost unchanged, resulting in no improvement in inter-laboratory CV for VL. This indicates that each in-house assay system worked well with good precision, but standardizing RM alone was insufficient for harmonization. The relevant choice of not only RM, but also internal control genes for data normalization is expected to be realistic to standardize HTLV-1 VL measurement. (Cancer Sci 2010; 101: 2361-2367).