生物靶标光敏剂对鸭乙肝病毒的特异结合及杀灭作用研究

RESEARCH ON SPECIFIC COMBINATION AND INACTIVATION OF BIO - TARGET PHOTOSENSITIVE AGENT ON DUCK HBV

目的:利用生物靶标效应的原理,探索一种特异灭活病毒的光化学方法。方法:将针对鸭乙肝病毒(DHBV)设计并人工合成的两条三螺旋结构寡核苷酸(TFO)分别标记8-甲氧基补骨脂素(8-MOP),并将这种复合物(TFO-P)与长波紫外线(UVA)共同作用于血液中的DHBV,通过原代细胞感染、酶联免疫测定、DMA斑点杂交等试验,观察在一定作用条件下,TFO-P对DHBV-DNA的特异结合及其所产生的特异光敏效应对DHBV的灭活程度。结果:DHBV-DNA样品斑点印迹随TFO-P含量的增加明显增强,而同时处理的血液中的白细胞DNA样品斑点印迹无明显增强;在TFO-P的加入量为0.1mmol/ml、UVA照射强度为180μw/cm^2时,5min～20min的UVA照射可灭活DHBV1.90～6.4个对数级,灭活病毒的效率显著高于UVA和8-MOP单独作用时的效果。结论:TFO-P与血液中DHBV-DNA能特异结合,在适宜的处理条件下,所产生的光敏效应能有效灭活血液中的DHBV,灭活滴度可达6个对数级以上。

Objective;To explore a photochemical method that can inactive hepatitis B virus(HBV)specifically based on the principle of bio - target.Method:Two synthesized TFOs designed to inactive duck HBV(DHBV)was marked with 8 - methoxypsoralen(8 - MOP), and then the combined effects of this compound(TFO - P)and UVA on DHBV in the blood were observed.The specific combination between TFO - P and DHBV - DNA and the degree of inactivation were determined by means of pre - generation infection, ELBA, dot -ELISA. Result:The blots of DHBV - DNA in samples were related directly to the density of TFO - P, while the WBC-DNA kept unchanged . The level of inactive DHBV could reach to 1.90-6.4 logarithm grades when the density of TFO - P 0.1 nmol/ml and UVA was 1800W/cm2.The combined inactive efficiency was higher significantly than they acted alone, respectively. Conclusion:This TFO- P can combined with DHBV - DNA in the blood specifically. The photosensitive agent can inactive DHBV in the blood with titer of over 6 logarithm grades under the appropriate conditions.