Detection of nucleic acid lesions during photochemical inactivation of RNA viruses by treatment with methylene blue and light using real-time PCR.
The mechanism of bacteriophage photoinactivation by methylene blue and light (MB+L) involves genomic RNA damage. In this study, two RNA viruses, Sindbis virus (SINV) and hepatitis C virus were treated by MB+L and their nucleic acids were amplified to show that RNA lesions occurred during inactivation. During MB+L inactivation, the viral load of both viruses was significantly reduced as MB+L exposure increased. The nucleic acid amplification of treated viral RNA was inhibited in a time-dependent manner and the percentage inhibition of amplification reached about 99% after 30 min of treatment. Furthermore, as compared to SINV viral infectivity detected by quantification of the 50% tissue culture infective dose (TCID(50)), the inhibition of SINV RNA amplification strongly correlated with a decrease in in vitro infectivity (R(2) > 0.94), suggesting that RNA serves as the main target during MB+L inactivation.
Shanghai Blood Center Graduate Student Training Base, East China Normal University, Shanghai, China.
Bo
B
Zhang
Lan
L
Zheng
Yuwen
Y
Huang
Qin
Q
Mo
Xun
X
Wang
Kaicheng
K
Qian
D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology; D002478:Cells, Cultured; D006224:Cricetinae; D008027:Light; D008751:Methylene Blue / Q000494:pharmacology*; D009696:Nucleic Acids / Q000737:chemistry* / Q000235:genetics; D016133:Polymerase Chain Reaction; D012327:RNA Virus Infections / Q000821:virology; D012328:RNA Viruses / Q000187:drug effects* / Q000528:radiation effects*
D000998:Antiviral Agents; D009696:Nucleic Acids; D008751:Methylene Blue