West Nile virus in plasma is highly sensitive to methylene blue-light treatment.
BACKGROUND
The epidemic of West Nile virus (WNV) in the US resulted in cases of transfusion-transmitted WNV. Effective pathogen reduction methods could have removed this infectious agent from the blood supply We have evaluated the efficacy of photodynamic treatment of fresh frozen plasma (FFP) with methylene blue (MB), a decontamination method applied in several European countries.


METHODS
FFP units (300 ml each) were spiked with WNV. MB was added, and the units were illuminated with white or monochromatic yellow light. WNV infectivity was determined by bioassay. WNV-RNA was quantitated by real-time PCR. The inactivation of WNV was investigated under standard and under suboptimal conditions, respectively. In addition, rechallenge experiments with multiple addition of WNV at maximal load (approx. 105 CFU/ml) and repeated illumination without replenishing MB were performed.


RESULTS
Complete inactivation of WNV was achieved by MB (0.8-1 mmol/l) and illumination with white light (30,000-45,000 Lux) within 2 min. White yellow light 20-40 J/cm(2) (2.5-5 min) were sufficient for inactivation by 5.75 log10-steps. The rechallenge experiments revealed the substantial reserve capacity of the procedure to inactivate WNV. Quantitative PCR indicated that the viral RNA was rapidly destroyed.


CONCLUSIONS
All experimental data demonstrate the enormous potency of phototreatment with MB to inactivate WNV in plasma.
Mohr
Harald
H
Blood Center of the German Red Cross Chapters of NSTOB, Institute Springe, Germany. hmohr@bsd-nstob.de
Knüver-Hopf
Joseph
J
Gravemann
Ute
U
Redecker-Klein
Anette
A
Müller
Thomas H
TH
D006801:Humans; D008027:Light; D008029:Lighting / Q000295:instrumentation; D008751:Methylene Blue / Q000494:pharmacology*; D010777:Photochemistry; D010949:Plasma / Q000821:virology*; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000187:drug effects / Q000528:radiation effects; D011838:Radiation-Sensitizing Agents / Q000494:pharmacology*; D038301:Virus Inactivation / Q000187:drug effects* / Q000528:radiation effects; D014902:West Nile virus / Q000187:drug effects* / Q000528:radiation effects
D012367:RNA, Viral; D011838:Radiation-Sensitizing Agents; D008751:Methylene Blue