Multilaboratory comparison of hepatitis C virus viral load assays
Multilaboratory comparison of hepatitis C virus viral load assays
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DOI：
10.1128/jcm.44.5.1726-1732.2006
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发表时间：
2006-05-01
影响因子：
9.4
通讯作者：
Lurain, N. S.
中科院分区：
文献类型：
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作者：
Caliendo, A. M.;Valsamakis, A.;Lurain, N. S.
我们报告了对丙型肝炎病毒 (HCV) 病毒载量测定的多实验室评估，以确定其线性范围、重现性、亚型检测和一致性。通过稀释临床样本（基因型 1b）构建一组名义浓度范围为 1.0 至 7.0 log(10) IU/ml 的 HCV RNA 样本。多个实验室使用 Abbott TaqMan 分析物特异性试剂（Abbott 逆转录 PCR [RT-PCR]）、Roche TaqMan RUO (Roche RT-PCR)、Roche Amplicor Monitor HCV 2.0 (Roche Monitor) 和 Bayer VERSANT HCV RNA 3.0 (Bayer bDNA) 检测对小组的重复进行了测试。使用拜耳 VERSANT HCV RNA 定性测定（拜耳 TMA）对拜耳 bDNA 阴性样本进行反射性测试。 Abbott RT-PCR 和 Roche RT-PCR 检测到浓度为 1.0 log,0 IU/ml 的所有 28 个重复，并且与 7.0 log(10) IU/ml 呈线性。罗氏监测仪和拜耳 bDNA 分别检测到 28 个重复中的 27 个和 28 个重复中的 13 个，浓度为 3.0 log,(, IU/ml。拜耳 TMA 检测到所有 7 个重复的浓度为 1.0 log(10) IU/ml。拜耳 bDNA 是四种检测中重复性最好的。在检测的线性范围内，小组成员的平均病毒载量值在不同的检测线性范围内为 0.5 log(10) 以内。测试。 在拜耳 bDNA、雅培 RT-PCR 和罗氏 RT-PCR 检测中对 89 个不同基因型（I 至 4）的临床样本进行了测试。对于 Abbott RT-PCR，基因型 1、2 或 3 样品的平均病毒载量值比拜耳 bDNA 测定值高 0.61 至 0.96 log(10)，基因型 4 的平均病毒载量值比拜耳 bDNA 测定值高 0.08 log(10) 标本。对于基因型 1、2 和 3 样本，罗氏 RT-PCR 测定得出的平均病毒载量值比拜耳 bDNA 测定所得的平均病毒载量值高 0.28 至 0.82 log(10)。然而，对于基因型 4 样品，通过罗氏 RT-PCR 测定获得的平均病毒载量值平均比拜耳 bDNA 低 0.15 log(10)。根据这些数据，我们 结论是 Abbott 和 Roche RT-PCR 检测的灵敏度和线性范围使其能够用于 HCV 诊断和治疗监测。然而，通过不同测定获得的病毒载量值的差异强调了在监测治疗反应时使用一种测定的重要性。
We report a multi laboratory evaluation of hepatitis C virus (HCV) viral load assays to determine their linear range, reproducibility, subtype detection, and agreement. A panel of HCV RNA samples ranging in nominal concentration from 1.0 to 7.0 log(10) IU/ml was constructed by diluting a clinical specimen (genotype 1b). Replicates of the panel were tested in multiple laboratories using the Abbott TaqMan analyte-specific reagent (Abbott reverse transcription-PCR [RT-PCR]), Roche TaqMan RUO (Roche RT-PCR), Roche Amplicor Monitor HCV 2.0 (Roche Monitor), and Bayer VERSANT HCV RNA 3.0 (Bayer bDNA) assays. Bayer bDNA-negative specimens were tested reflexively using the Bayer VERSANT HCV RNA qualitative assay (Bayer TMA). Abbott RT-PCR and Roche RT-PCR detected all 28 replicates with a concentration of 1.0 log,0 IU/ml and were linear to 7.0 log(10) IU/ml. Roche Monitor and Bayer bDNA detected 27 out of 28 and 13 out of 28 replicates, respectively, of 3.0 log,(, IU/ml. Bayer TMA detected all seven replicates with 1.0 log(10) IU/ml. Bayer bDNA was the most reproducible of the four assays. The mean viral load values for panel members in the linear ranges of the assays were within 0.5 log(10) for the different tests. Eighty-nine clinical specimens of various genotypes (I through 4) were tested in the Bayer bDNA, Abbott RT-PCR, and Roche RT-PCR assays. For Abbott RT-PCR, mean viral load values were 0.61 to 0.96 log(10) greater than the values for Bayer bDNA assay for samples with genotype 1, 2, or 3 samples and 0.08 log(10) greater for genotype 4 specimens. The Roche RT-PCR assay gave mean viral load values that were 0.28 to 0.82 log(10) greater than those obtained with the Bayer bDNA assay for genotype 1, 2, and 3 samples. However, for genotype 4 samples the mean viral load value obtained with the Roche RT-PCR assay was, on average, 0.15 log(10) lower than that of the Bayer bDNA. Based on these data, we conclude that the sensitivity and linear range of the Abbott and Roche RT-PCR assays enable them to be used for HCV diagnostics and therapeutic monitoring. However, the differences in the viral load values obtained with the different assays underscore the importance of using one assay when monitoring response to therapy.