Human Protein Process Sciences, Lille, France.
Vox Sang. 2013 Jan;104(1):1-6. doi: 10.1111/j.1423-0410.2012.01621.x. Epub 2012 Jul 4.
A minipool solvent/detergent (S/D; 1% TnBP/1% Triton X-45; 31°C) process was developed for viral inactivation of plasma and cryoprecipitate used for transfusion. The goal of this study was to determine the rate and extent of inactivation of dengue virus (DENV) during this process.
DENV-1 was propagated using C6/36 mosquito cells to an infectivity titre close to 9 log and spiked (10% v/v) into individual plasma and cryoprecipitate samples from two distinct donors. Samples were taken right after spiking and during viral inactivation treatment by 1% TnBP-1% Triton X-45 at 31°C. DENV-1 infectivity was assessed on Vero E6 cells by a focus-forming assay (FFA). Culture medium and complement-inactivated plasma were used as experimental controls. Experiments were done in duplicate.
DENV-1 infectivity was 7·5 log in spiked plasma and 7·1 and 7·3 log in spiked cryoprecipitate. There was no loss of DENV-1 infectivity in the spiked materials, nor in the controls not subjected to S/D treatment. No infectivity was found in plasma and cryoprecipitate subjected to S/D treatment at the first time-point evaluated (10 min).
DENV-1 was strongly inactivated in plasma and cryoprecipitate, respectively, within 10 min of 1% TnBP/1% Triton X-45 treatment at 31°C. These data provide a reassurance of the safety of such S/D-treated plasma and cryoprecipitate with regard to the risk of transmission of all DENV serotypes and other flaviviruses.
已开发出一种用于病毒灭活的小型混合溶剂/去污剂(S/D;1%TnBP/1% Triton X-45;31°C)工艺,用于处理血浆和冷沉淀以进行输血。本研究的目的是确定在该过程中登革热病毒(DENV)的失活动力速率和程度。
使用 C6/36 蚊细胞培养 DENV-1 至接近 9 对数级的感染滴度,并将其分别掺入来自两个不同供体的单个血浆和冷沉淀样本中(10%v/v)。在病毒失活处理开始时(31°C 下用 1%TnBP-1% Triton X-45 处理)和处理过程中,立即从加入病毒的样本中取样。通过噬斑形成测定(FFA)评估 Vero E6 细胞中的 DENV-1 感染力。使用培养基和补体失活的血浆作为实验对照。实验重复进行两次。
掺入血浆中的 DENV-1 感染力为 7.5 对数级,掺入冷沉淀中的感染力为 7.1 和 7.3 对数级。在未进行 S/D 处理的掺入材料和对照中,DENV-1 感染力均未丧失。在首次评估的时间点(10 分钟),未在接受 S/D 处理的血浆和冷沉淀中发现感染力。
在 31°C 下用 1%TnBP/1% Triton X-45 处理 10 分钟内,DENV-1 在血浆和冷沉淀中分别被强烈灭活。这些数据为 S/D 处理的血浆和冷沉淀的安全性提供了保证,可降低所有 DENV 血清型和其他黄病毒的传播风险。