Duck hepatitis B photoinactivation bydimethylmethylene blue in RBC suspensions.
BACKGROUND
Dimethylmethylene blue (DMMB) has been used to photoinactivate a number of model viruses, including VSV, in RBC suspensions under conditions that preserve in vitro RBC properties during storage. The relative sensitivity of duck HBV (DHBV) and VSV to photoinactivation by DMMB was investigated by performing an indirect immunofluorescence assay (IFA) using primary duck hepatocyte (PDH) cultures or a standard plaque assay for the respective viruses.


METHODS
DMMB was added to 45-percent Hct, WBC-reduced, oxygenated AS-3 RBCs at 10-, 1-, and 0.1-microM concentrations. Samples (1-mm thick) were illuminated with 5.4-mW per cm(2) of red light for 2 or 9 seconds. Unilluminated samples without DMMB or with 10 microM DMMB served as control.


RESULTS
DHBV and VSV were rapidly photoinactivated by DMMB in a concentration and light-dose-dependent fashion. Neither virus was substantially inactivated by incubation with DMMB in the dark. For a given light exposure, DHBV required a concentration of DMMB one-one hundredth that of VSV to achieve approximately the same level of inactivation.


CONCLUSIONS
DHBV appears to be considerably more sensitive than VSV to DMMB photoinactivation. Photoinactivation in 45-percent Hct RBCs can be achieved in seconds by using micromolar quantities of dye.
Wagner
S J
SJ
American Red Cross Holland Laboratory for the Biomedical Sciences, Rockville, Maryland, USA. wagners@usa.redcross.org
Skripchenko
A
A
Pugh
J C
JC
Suchmann
D B
DB
Ijaz
M K
MK
D004305:Dose-Response Relationship, Drug; D004307:Dose-Response Relationship, Radiation; D004912:Erythrocytes / Q000821:virology*; D019084:Fluorescent Antibody Technique, Indirect; D015611:Hepatitis B Virus, Duck / Q000187:drug effects* / Q000528:radiation effects*; D006801:Humans; D008027:Light; D008751:Methylene Blue / Q000031:analogs & derivatives / Q000494:pharmacology*; D014775:Virus Activation / Q000187:drug effects*
C435946:dimethylmethylene blue; D008751:Methylene Blue