Effect of ultrasound with methylene blue as sound sensitive agent on virus inactivation

The goal of this paper is to explore a method for virus inactivation based on ultrasonic treatment, and on this basis, to explore the synergistic effect of methylene blue as a sonosensitizer in virus inactivation. The titer of human parainfluenza virus type 3 (HPIV3), Autographacaliforinica nuclear polyhedrosis virus carried a green fluorescent reporter gene (AcNPV-GFP) and Enterovirus group D 68 (EV-D68) were determined by plaque assays or TCID50 methods after treatment by MB combined with illumination (MB photochemical, MBP) or ultrasonic excitation. Different ultrasonic power and time, MB concentration gradient were set to determine the best anti-viral combination. For the enveloped virus HPIV3 and AcNPV-GFP, pure ultrasonic or MBP treatment could reduce viral titer more than 104，and ultrasonic combined with MB could completely inactivate HPIV3 and AcNPV-GFP in 5min while traditional MBP methods could only reduce viral titer about 10X. For the non-enveloped virus EV-D68, pure ultrasonic or MBP treatment only reduced the viral titer about 102, but ultrasonic combined with MB treatment could reduce the viral titer about 103 in 5min and completely inactive EV-D68 in 10min (reduced 105). Compared with MBP method and pure ultrasonic inactivation, ultrasonic combined with MB has better inactivation effect on either enveloped or non-enveloped viruses, and the appropriate combination of parameters is expected to be a new blood transfusion transmitted virus inactivation method.