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Merck

Dengue virus inactivation by minipool TnBP/Triton X-45 treatment of plasma and cryoprecipitate.

Vox sanguinis (2012-07-05)
T Burnouf, M-L Chou, L-H Cheng, Z-R Li, Y-W Wu, M El-Ekiaby, K-H Tsai
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A minipool solvent/detergent (S/D; 1% TnBP/1% Triton X-45; 31ยฐC) 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 31ยฐC. 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 7ยท5 log in spiked plasma and 7ยท1 and 7ยท3 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 31ยฐC. 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.

MATERIALS
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Sigma-Aldrich
Tributyl phosphate, ≥99%
Supelco
Tributyl phosphate, for extraction analysis, ≥99.0% (GC)
Sigma-Aldrich
Tributyl phosphate, 97%
Supelco
Tributyl phosphate, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
Tributyl phosphate, puriss., ≥99.0% (GC)
Tri-n-butyl phosphate, European Pharmacopoeia (EP) Reference Standard