Boujnan Mohamed, Duits Ashley J, Koppelman Marco H G M
Sanquin Blood Supply Foundation, National Screening Laboratory of Sanquin (NSS), Amsterdam, the Netherlands.
Biomedical & Health Research Institute, Willemstad, Curaçao.
Transfusion. 2018 Mar;58(3):641-648. doi: 10.1111/trf.14460. Epub 2018 Jan 10.
Several countries have implemented safety strategies to reduce the risk of Zika virus (ZIKV) transmission through blood transfusion. These strategies have included nucleic acid amplification testing (NAT) of blood donations. In this study, a new real-time polymerase chain reaction (PCR) assay including internal control for the detection of ZIKV on the cobas omni Utility Channel (UC) on the cobas 6800 system is presented.
PCR conditions and primer/probe concentrations were optimized on the LightCycler 480 instrument. Optimized conditions were transferred to the cobas omni UC on the cobas 6800 system. Subsequently, the limit of detection (LOD) in plasma and urine, genotype inclusivity, specificity, cross-reactivity, and clinical sensitivity were determined.
The 95% LOD of the ZIKV PCR assay on the cobas 6800 system was 23.0 IU/mL (95% confidence interval [CI], 16.5-37.5) in plasma and 24.5 IU/mL (95% CI, 13.4-92.9) in urine. The assay detected African and Asian lineages of ZIKV. The specificity was 100%. The clinical concordance between the newly developed ZIKV PCR assay and the investigational Roche cobas Zika NAT test was 83% (24/29).
We developed a sensitive ZIKV PCR assay on the cobas omni UC on the cobas 6800 system. The assay can be used for large-scale screening of blood donations for ZIKV or for testing of blood donors returning from areas with ZIKV to avoid temporal deferral. This study also demonstrates that the cobas omni UC on the cobas 6800 system can be used for in-house-developed PCR assays.
多个国家已实施安全策略以降低通过输血传播寨卡病毒（ZIKV）的风险。这些策略包括对献血进行核酸扩增检测（NAT）。在本研究中，我们展示了一种新的实时聚合酶链反应（PCR）检测方法，该方法包括用于在cobas 6800系统的cobas omni通用通道（UC）上检测ZIKV的内部对照。
在LightCycler 480仪器上优化了PCR条件以及引物/探针浓度。将优化后的条件转移至cobas 6800系统的cobas omni UC上。随后，测定了血浆和尿液中的检测限（LOD）、基因型包容性、特异性、交叉反应性以及临床敏感性。
cobas 6800系统上ZIKV PCR检测方法的95% LOD在血浆中为23.0 IU/mL（95%置信区间[CI]，16.5 - 37.5），在尿液中为24.5 IU/mL（95% CI，13.4 - 92.9）。该检测方法可检测ZIKV的非洲和亚洲谱系。特异性为100%。新开发的ZIKV PCR检测方法与罗氏cobas寨卡病毒NAT检测试验之间的临床一致性为83%（24/29）。
我们在cobas 6800系统的cobas omni UC上开发了一种灵敏的ZIKV PCR检测方法。该检测方法可用于对献血进行ZIKV的大规模筛查，或用于检测从有ZIKV地区返回的献血者，以避免暂时延期。本研究还表明，cobas 6800系统上的cobas omni UC可用于内部开发的PCR检测方法。
