摘要
为观察特异光敏剂对红细胞悬液中细胞内DHBV的抑制作用,探索全血病毒灭活方法。将设计、合成末端标记光敏剂的三螺旋结构寡核苷酸序列(TFO-P),以适当剂量加入模拟的病毒污染血液,在UVA(长波紫外线)的协同下处理血液;以PCR技术检测细胞培养的上清液中的DHBV-DNA,以组织原位杂交技术检测原代细胞内的DHBV,观察被处理前后肝原代细胞内DHBV的活性。结果,在经0.1nmol/ml TFO-P和1800μW/cm^2UVA照射10min后,分离、培养的鸭肝细胞上清液内检不出DHBV-DNA,细胞内DHBV-DNA在第7日转阴,而对照组的培养液和细胞内均可检出DHBV-DNA。结论,试验所设计的特异光敏剂能有效抑制血细胞悬液中细胞内的DHBV。
In order to explore a new technique to inactive the virus in blood, the efficacy of a photosensitizer having the targeting effect in inhibiting intracelular DHBV was studied. A photosensitizer (TFO-P) targeting DHBV-DNA X/ C region, a specific sequence decorated by photosensitizer at its end, was designed and synthesized. Then the TFO-P was added to the contaminated blood, which was irradiated with long wave ultraviolet ray (UVA) for a designed time, the DHBV-DNA in supernatant of cultures and the DHBV activity in primaty duck hepatocytes (PDH) were detected by PCR assay and in situ hybridization respectively. When the added dose of TFO-P was about 0.1 nmol/ ml, and then irradiated by UVA for 10 min at the intensity of 1 800 μW/ cm2, the DHBA-DNA in the supernatant of culture was not detected at all. The DHBV-DNA in PDH became negative on seventh day. In the control group, the DHBV-DNA could be detected at all observing time. Conclusion: The specially designed photosensitizer could inhibit the intracellular DHBV.
出处
《中国消毒学杂志》
CAS
2004年第1期24-26,共3页
Chinese Journal of Disinfection
基金
国家自然科学基金(39870693)
全军九五科技课题基金(98M096)
关键词
光敏剂
三螺旋结构
寡核苷酸
DHBV
抑制
photo sensitizer, Triple helix form
oligonucleotide
DHBV
Inactivation
