Sutherland Michael R, Simon Ayo Y, Serrano Katherine, Schubert Peter, Acker Jason P, Pryzdial Edward L G
Centre for Innovation, Canadian Blood Services, Vancouver, British Columbia, Canada.
Centre for Blood Research and Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, British Columbia, Canada.
Transfusion. 2016 May;56(5):1129-37. doi: 10.1111/trf.13454. Epub 2016 Jan 18.
Dengue virus (DENV) is a transfusion-transmissible arbovirus that threatens blood donor systems with approximately 200 million high-titer asymptomatic infections occurring annually. Here we investigated the viability of DENV during storage of donor-derived platelet (PLT) and red blood cell (RBC) units. While purified PLTs have been shown to generate viable DENV, RBCs are replication incompetent. Combined with different storage criteria, distinct virus persistence profiles were anticipated in PLT and RBC units.
Mimicking the virus titer of asymptomatic donors, purified DENV was spiked (10(5) -10(6) infectious units/mL) into PLT or RBC units produced and stored according to blood bank operating procedures. DENV was measured by infectious plaque-forming assays and by quantitative reverse transcription-polymerase chain reaction.
In both PLT (7 days, 20-24°C) and RBC (42 days, 1-6°C) units, infectious DENV persisted throughout storage despite logarithmic decay. In buffer alone, DENV infectivity was insignificant by Day 1 at 20 to 24°C or 14 days at 1 to 6°C. Infectious virus production was identified in stored PLT units using a translation inhibitor and supported by virus genome replication. Surprisingly, DENV was also produced in RBC units, implying the involvement of cells other than RBCs.
Both virus propagation and effects independent of cell function mitigate the intrinsic lability of DENV. Nevertheless, the overall rapid storage decay suggests that aged PLT and RBC units may be safer. These data raise awareness to the possible persistence of other conceivably more robust RNA viruses during the storage of cellular blood products.
登革病毒(DENV)是一种可通过输血传播的虫媒病毒,每年约有2亿例高滴度无症状感染病例,对献血者系统构成威胁。在此,我们研究了登革病毒在供体来源的血小板(PLT)和红细胞(RBC)单位储存期间的生存能力。虽然已证明纯化的血小板可产生有活力的登革病毒,但红细胞无复制能力。结合不同的储存标准,预计血小板和红细胞单位中病毒的持续存在情况会有所不同。
模仿无症状献血者的病毒滴度,将纯化的登革病毒(10⁵ - 10⁶感染单位/毫升)加入按照血库操作程序生产和储存的血小板或红细胞单位中。通过感染性空斑形成试验和定量逆转录 - 聚合酶链反应检测登革病毒。
在血小板单位(7天,20 - 24°C)和红细胞单位(42天,1 - 6°C)中,尽管病毒呈对数衰减,但整个储存期间仍存在有感染性的登革病毒。仅在缓冲液中,登革病毒在20至24°C下第1天或1至6°C下第14天时感染性就微不足道了。使用翻译抑制剂在储存的血小板单位中鉴定出有感染性的病毒产生,并得到病毒基因组复制的支持。令人惊讶的是,红细胞单位中也产生了登革病毒,这意味着除红细胞外还有其他细胞参与。
病毒传播和与细胞功能无关的效应均减轻了登革病毒固有的不稳定性。然而,整体快速的储存衰减表明,陈旧的血小板和红细胞单位可能更安全。这些数据提高了人们对在细胞血液制品储存期间其他可能更稳定的RNA病毒可能持续存在的认识。