Mah Jordan, Huang Chun Hong, Sahoo Malaya K, Pinsky Benjamin A
Department of Pathology, Stanford University School of Medicine, Stanford, California, USA.
Department of Medicine, Division of Infectious Diseases and Geographic Medicine, Stanford University School of Medicine, Stanford, California, USA.
Microbiol Spectr. 2023 Feb 27;11(2):e0501022. doi: 10.1128/spectrum.05010-22.
Human adenoviruses (HAdVs) cause severe disease in immunocompromised patients. Quantitation of HAdV DNA in peripheral blood is used to assess the risk of disseminated disease and to monitor response to therapy. The lower limit of detection, precision, and linearity of the semiautomated AltoStar adenovirus quantitative PCR (qPCR) was evaluated using reference HAdV-E4 in EDTA plasma and respiratory virus matrix. Qualitative and quantitative agreement was determined using 122 clinical EDTA plasma specimens previously tested using a laboratory-developed HAdV qPCR. The 95% lower limit of detection (LLOD) was 33 IU/mL (95% confidence interval [CI], 10 to 56) for EDTA plasma and 188 IU/mL (95% CI, 145 to 304) for respiratory swab matrix. In both matrices, the AltoStar HAdV qPCR was linear from 7.0 to 2.0 log IU/mL. For the clinical specimens, overall agreement was 96.7% (95% CI, 91.8 to 99.1), positive percent agreement was 95.5% (95% CI, 87.6 to 98.5), and negative percent agreement was 98.2% (95% CI, 88.5 to 99.7). Passing-Bablok analysis of specimens quantifiable by both methods revealed a regression line of Y = 1.11 · X + 0.00; there was positive proportional bias (95% CI of the slope, 1.05 to 1.22) but no systematic bias (95% CI of the Y-intercept, -0.43 to 0.23) compared to the reference. The AltoStar platform provides accurate quantitation of HAdV DNA and provides a semiautomated option for the clinical monitoring of HAdV following transplantation. Accurate quantification of human adenovirus DNA in the peripheral blood plays a critical role in the management of adenovirus infections in transplant recipients. Many laboratories utilize in-house laboratory-based PCR assays for the quantification of human adenovirus, as there are few commercial options available. Here, we describe the analytical and clinical performance of the semiautomated AltoStar adenovirus quantitative PCR (Altona Diagnostics). This platform provides sensitive, precise, and accurate quantification of adenovirus DNA that is well suited for virological testing following transplantation. Prior to implementing a new quantitative test in the clinical laboratory, a rigorous evaluation is required to determine assay performance characteristics and to correlate results to current in-house methods of quantitation.
人腺病毒（HAdVs）可导致免疫功能低下患者发生严重疾病。对外周血中的HAdV DNA进行定量分析，可用于评估播散性疾病的风险以及监测治疗反应。使用参考HAdV-E4，在乙二胺四乙酸（EDTA）血浆和呼吸道病毒基质中评估了半自动AltoStar腺病毒定量聚合酶链反应（qPCR）的检测下限、精密度和线性度。通过使用122份先前采用实验室自行研发的HAdV qPCR检测过的临床EDTA血浆标本，确定了定性和定量的一致性。对于EDTA血浆，95%检测下限（LLOD）为33 IU/mL（95%置信区间[CI]，10至56）；对于呼吸道拭子基质，95%检测下限为188 IU/mL（95% CI，145至304）。在两种基质中，AltoStar HAdV qPCR在7.0至2.0 log IU/mL范围内呈线性。对于临床标本，总体一致性为96.7%（95% CI，91.8至99.1），阳性百分比一致性为95.5%（95% CI，87.6至98.5），阴性百分比一致性为98.2%（95% CI，88.5至99.7）。对两种方法均可定量的标本进行的Passing-Bablok分析显示，回归线为Y = 1.11·X + 0.00；与参考方法相比，存在正比例偏差（斜率的95% CI，1.05至1.22），但无系统偏差（Y轴截距的95% CI，-0.43至0.23）。AltoStar平台可准确定量HAdV DNA，并为移植后HAdV的临床监测提供了半自动选项。对外周血中的人腺病毒DNA进行准确量化，在移植受者腺病毒感染的管理中起着关键作用。由于可供选择的商业检测方法很少，许多实验室使用基于实验室内部的PCR检测方法来定量人腺病毒。在此，我们描述了半自动AltoStar腺病毒定量PCR（Altona Diagnostics公司）的分析性能和临床性能。该平台可对腺病毒DNA进行灵敏、精确且准确的定量，非常适合移植后的病毒学检测。在临床实验室实施一项新的定量检测之前，需要进行严格评估，以确定检测性能特征，并将结果与当前的实验室内部定量方法进行关联。
