Vitalant Research Institute, San Francisco, California, United States of America.
Department of Laboratory Medicine, University of California San Francisco, San Francisco, California, United States of America.
PLoS Negl Trop Dis. 2023 Mar 17;17(3):e0011157. doi: 10.1371/journal.pntd.0011157. eCollection 2023 Mar.
Early detection of Zika virus (ZIKV) transmission within geographic regions informs implementation of community mitigation measures such as vector reduction strategies, travel advisories, enhanced surveillance among pregnant women, and possible implementation of blood and organ donor screening or deferral. Standardized, comparative assessments of ZIKV assay and testing lab performance are important to develop optimal approaches to ZIKV diagnostic testing and surveillance. We conducted an expanded blinded panel study to characterize and compare the analytical performance of fifteen diagnostic and blood screening ZIKV NAT assays, including detection among single- and multiplex assays detecting ZIKV, dengue virus (DENV) and chikungunya virus (CHIKV). A 300 member blinded panel was constructed, consisting of 11 serial half-log dilutions ranging from ~104 to 10-1 genome equivalents/mL in 25 replicates each of the Tahitian Asian ZIKV isolate in ZIKV-negative human serum. Additionally, clinical samples from individuals with DENV-like syndrome or suspected ZIKV infection in Brazil were evaluated. The majority of assays demonstrated good specificity. Analytical sensitivities varied 1-2 logs, with a substantially higher limit of detection (LOD) in one outlier. Similar analytical sensitivity for ZIKV RNA detection in singleplex and multiplex assays of the Grifols and ThermoFisher tests were observed. Coefficient of Assay Efficiency (CE), calculated to characterize assays' RNA extraction and amplification efficiency, ranged from 0.13 for the Certest VIASURE multiplex and 0.75 for the Grifols multiplex assays. In general, assays using transcription mediated amplification (TMA) technology had greater CE compared to assays using conventional PCR technology. Donor screening NAT assays were significantly more sensitive than diagnostic RT-qPCR assays, primarily attributable to higher sample input volumes. However, ideal assays to maximize sensitivity and throughput may not be a viable option in all contexts, with other factors such as cost, instrumentation, and regulatory approval status influencing assay availability and selection, particularly in resource constrained settings.
早期检测地理区域内的寨卡病毒（ZIKV）传播情况，可为实施社区缓解措施提供信息，如减少病媒策略、旅行建议、加强孕妇监测以及可能实施血液和器官供者筛查或延迟。对寨卡病毒检测和检测实验室性能进行标准化、比较评估，对于制定最佳寨卡病毒诊断检测和监测方法非常重要。我们进行了一项扩展的盲法检测板研究，以描述和比较 15 种诊断和血液筛查寨卡病毒 NAT 检测方法的分析性能，包括在检测寨卡病毒、登革热病毒（DENV）和基孔肯雅热病毒（CHIKV）的单重和多重检测中。构建了一个由 300 名成员组成的盲法检测板，该检测板由 11 个连续半对数稀释度组成，范围从每毫升约 104 到 10-1 基因组当量/ml，每个稀释度重复 25 次，在 ZIKV 阴性人血清中的 Tahitian Asian ZIKV 分离株中。此外，还评估了来自巴西具有登革热样综合征或疑似寨卡病毒感染的临床样本。大多数检测方法表现出良好的特异性。分析灵敏度相差 1-2 个对数，一个异常值的检测限（LOD）明显较高。在格里福斯和赛默飞世尔的检测中，单重和多重检测中寨卡病毒 RNA 的检测分析灵敏度相似。为了描述检测的 RNA 提取和扩增效率，计算了检测效率系数（CE），范围从 Certest VIASURE 多重检测的 0.13 到格里福斯多重检测的 0.75。一般来说，使用转录介导扩增（TMA）技术的检测方法比使用传统 PCR 技术的检测方法具有更高的 CE。供者筛查 NAT 检测方法比诊断 RT-qPCR 检测方法更灵敏，这主要归因于更高的样本输入量。然而，在所有情况下，并非所有情况下都能选择到理想的检测方法以最大限度地提高灵敏度和通量，其他因素如成本、仪器和监管批准状态会影响检测的可用性和选择，特别是在资源有限的环境中。
