Evaluation of the Virucidal Activity of Chlorine Dioxide against Hepatitis B Virus

Hepatitis B virus (HBV) remains a major global public health concern, with numerous nosocomial and occupational outbreaks reported. Preventing indirect HBV transmission through effective disinfection is crucial. This study evaluated the virucidal activity of chlorine dioxide (ClO₂) against HBV and compared it with sodium hypochlorite (NaClO). Pretreatment of HBV with ClO₂ at 10, 100, and 250 ppm (mg/L) for 1 min reduced hepatitis B surface antigen (HBsAg) release to 65.5%, 21.9%, and 7.0%; hepatitis B core (HBc)-positive cells to 59.1%, 27.0%, and 6.6%; intracellular HBV RNA to 71.5%, 31.6%, and 3.0%; and extracellular HBV DNA to 82.2%, 42.6%, and 14.1%, respectively. At identical concentrations, NaClO produced smaller reductions. In the presence of organic matter, no significant reduction in ClO₂ activity against HBV was observed. ClO₂ demonstrated approximately 2.2-fold greater anti-HBV potency than NaClO with and without bovine serum albumin and sheep erythrocytes. Neutralizer, cell-permissiveness, and ultrafiltration controls supported direct viral inactivation. These findings indicate that ClO₂ showed stronger in vitro anti-HBV activity than NaClO within the tested concentration range and conditions.