Comparison of different HCV viral load and genotyping assays
Comparison of different HCV viral load and genotyping assays
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DOI：
10.1016/s1386-6532(02)00235-4
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发表时间：
2003-09-01
影响因子：
8.8
通讯作者：
Williams, KJ
中科院分区：
文献类型：
--
作者：
Anderson, JC;Simonetti, J;Williams, KJ
背景资料：我们报告了一个实验室间比较的方法，用于确定丙型肝炎病毒（HCV）血清载量和基因型之间的最近成立的分子实验室在阿拉斯加土著医学中心（ANMC）和两个独立的实验室使用不同的检测。在ANMC，已经开发了实时定量RT-PCR扩增方法（QPCR），其中通过将QPCR结果内插到根据世界卫生组织（WHO）第一个HCV国际标准品校准的标准品的结果来确定HCV病毒载量。随后通过对QPCR检测产生的DNA片段进行直接测序来确定HCV基因型。目的和研究设计：将上述方法与由两个独立实验室使用不同的市售病毒载量测定法，Quantiplex（TM）HCV RNA（Bayer Diagnostics）和Amplicor（TM）HCV Monitor（TM）（v2.0）（Roche Molecular Systems），以及两种不同的基因分型测定法，限制性片段长度多态性（RFLP）和INNO-LiPA HCV 11（Innogenetics），对相同患者血清获得的结果进行统计学比较。结果如下：ANMC的实时QPCR HCV病毒载量结果与Quantiplex（TM）HCV RNA方法所得结果相当（R-2 = 0.3813），并且与最近批号的Amplicor（TM）HCV Monitor（TM）（其中病毒载量以IU/ml导出）相当好地比较（R-2 = 0.6408），但与早期批号的Amplicor（TM）HCV Monitor（TM）相比较差，其中病毒载量以拷贝/ml为单位（R-2 = 0.0913）。用于基因型测定的ANMC直接测序方法与RFLP（84-86%）和INNO-LiPA（85-97.5%）方法进行了中等至非常好的比较。结论：这些病毒载量比较突出了使用不同类型的检测试剂监测患者HCV病毒载量时可能发生的差异。用不同方法比较HCV基因型在统计学上更可靠，对预测抗病毒治疗的应答率有重要的临床意义。然而，一旦治疗开始，病毒载量对于监测反应是重要的。(C)2002 Elsevier Science B. V.保留所有权利。
Background: We report an interlaboratory comparison of methods for the determination of hepatitis C virus (HCV) serum load and genotype between a recently, established molecular laboratory at the Alaska Native Medical Center (ANMC) and two independent laboratories using different assays. At ANMC, a Real-time quantitative RT-PCR amplification methodology (QPCR) has been developed in which HCV viral loads are determined by interpolation of QPCR results to those of standards calibrated to the World Health Organization (WHO) First International Standard for HCV. HCV genotype is subsequently determined by direct sequencing of the DNA fragment generated from the QPCR assay. Objectives and Study Design: The above methods were statistically compared to results obtained for the same patient sera by two independent laboratories using different commercially available viral load assays; Quantiplex(TM) HCV RNA (Bayer Diagnostics) and Amplicor(TM) HCV Monitor(TM) (v 2.0) (Roche Molecular Systems), as well as two different genotyping assays; restriction fragment length polymorphism, (RFLP) and INNO-LiPA HCV 11 (Innogenetics). Results: ANMC's Real-time QPCR HCV viral load results compared moderately well with those obtained by the Quantiplex(TM) HCV RNA method (R-2 = 0.3813), and compared quite well with recent lot numbers of Amplicor(TM) HCV Monitor(TM) in which viral loads are derived in IU/ml (R-2 = 0.6408), but compared poorly with earlier lot numbers of Amplicor(TM) HCV Monitor(TM) in which viral loads were derived in copies/ml (R-2 = 0.0913). The ANMC direct sequencing method for genotype determination compared moderately to very well with both the RFLP (84-86%) and INNO-LiPA (85-97.5%) methods. Conclusions: These viral load comparisons highlight the discrepancies that may occur when patient HCV viral loads are monitored using different types of assays. Comparison of HCV genotype by different methods is more reliable statistically and important clinically for predicting probability of response to antiviral therapy. However, viral loads are important for monitoring response once therapy has begun. (C) 2002 Elsevier Science B.V. All rights reserved.