目的 利用鸭乙型肝炎病毒(DHBV)感染动物模型,评价亚甲蓝光化学病毒灭活方法对血液成分中DNA病毒的灭活效果。方法 将超离纯化的DHBV分别加入人血浆或人红细胞,经亚甲蓝光化学灭活病毒,将含不同基因组拷贝数DHBV的血浆成分经静脉感染1 d龄雏鸭。采用放射性核素核酸杂交法对血清中DHBV DNA进行检测,计算病毒灭活处理前、后人血浆及人红细胞中DHBV的半数感染计量(ID50)。结果 结果显示加入DHBV的血浆在未经灭活处理前对1 d龄雏鸭的ID50值为103.33,而经病毒灭活处理后ID50值为1010拷贝,灭活处理可使病毒感染性滴度下降达6个Log;加入DHBV的红细胞灭活前ID50值为103.35,经灭活处理后ID50值为108.35拷贝,灭活处理使病毒感染性滴度下降5个Log。结论 利用DHBV感染动物模型,可以检测到少量病毒在自然感染宿主体内的感染性,可用于评判血液成分中病毒灭活方法的效果,亚甲蓝光化学处理对血浆中DNA病毒的灭活效果较好于对红细胞中DNA病毒的灭活作用。

Objective Duck hepatitis B virus (DHBV) infected animal model was used to evaluate the inactivation of DNA virus by methylene blue photodynamic method. Methods DHBV was purified by Ultra-centrifugation. Purified DHBV was sequentially diluted and added to human plasma or RBC components respectively, followed by inactivation with methylene blue photodynamic method. These samples were inactivated into one-day-old ducklings by intravenous injection. Sera from ducks were collected at different intervals and detected by dot hybridization with DHBV probe labeled with 32P for DHBV DNA, and confirmed by southern blot hybridization for DHBV DNA in duck livers. Results The infectivity of DHBV (ID50) in the blood components was calculated before and after the treatment with methylene blue photodynamic method. Before viral inactivation, the ID50 of DHBV mixed with human plasma for one-day-old duckling was 1033.3 copies, and the ID50 of DHBV mixed with human RBC in one-day-old duckling was 103.50 copies. After inactivation with methylene blue photodynamic, the ID50 of DHBV mixed with plasma in one-day-old duckling was 1010 copies, while the ID50 of DHBV-RBC in one-day-old duckling was 108.35 copies. Treatement of methylene blue photodynamic the viral titer of DHBV-plasma decreased 6 logs, and DHBV-RBC decreased 5 logs. Conclusion Methylene blue photodynamic can inactivate DNA virus(using DHBV as a marker) in human blood or RBC, and the inactivation effect is better for virus in the plasma than that in RBC. The DHBV infected ducks could be used as a model for evaluating the effect of DNA virus inactivation in vivo.