特异光敏效应对血细胞悬液中DHBV的灭活作用

The inactivation of duck hepatitis virus B in blood cell suspenion induced by specific photosensitizer

目的 观察设计的靶向光敏剂所产生的特异光化学效应对血细胞悬液中病毒的灭活作用 ,以探索新型的血液病毒灭活技术。方法 以光化学效应为基本原理 ,利用反义核酸技术设计合成与DHBV DNA特异结合的TFO P ,将其作为特异的靶向光敏剂 ,在UVA的协同下将其作用于血细胞悬液中的DHBV。通过电泳转移印迹试验、原代鸭肝细胞感染试验 ,观察TFO P与DHBV DNA的结合效应和在UVA协同下灭活DHBV的效果。结果 所设计的TFO P能与不同株DHBV DNA样本产生不同程度的特异结合 ,在剂量为 0 .1nmol/ml的TFO P和照射强度为 180 0 μW/cm2 的UVA作用下 ,血细胞悬液中DHBV的活性下降 1.9～ 5 .4Log。 结论 TFO P所产生的光化学效应能显著灭活血细胞悬液中的DHBV。

Objective:To study the specific photochemical effects of a newly designed target photosensitizer. Methods Based on the technique of antisense nucleic acid and the principle of photochemical reaction effects,a specific sensitizer,TFO P has been designed and synthesized.When in coordination with long wave ultraviolet ray(UVA) ,this decorated complex (TFO P) was added into the blood cell suspension to inactivate the contaminating virus( duck hepatitis virus B,DHBV).The efficacy of specific binding to DHBV DNA and viral inactivation by TFO P was detected by gel shift blot assay and infection of primary culture of duck hepatocyte.Results The designed TFO P could specifically bind to different DHBV DNA line sample and present different linking level.With a TFO P concentration of 0.1 nmol/ml and UVA intensity of 1800 μW/cm 2,the DHBV in blood cell suspension could be reduced by 1.90～5.40 logs.Conclcusion The photochemical effects of TFO P could significant inactivate DHBV in blood.