核黄素光化学作用灭活大肠杆菌的研究

目的研究核黄素光化学反应对细菌的灭活作用,探讨分析核黄素光化学灭活病原体的机理。方法将含有细菌的培养液4.950ml注入5ml血袋中,再加入10mmol/L的核黄素溶液50μl使其终浓度为100μmol/L,将血袋置控温光照仪中接受400—500nm波段可见光双侧照射剂量(8.0—32.0)J/cm2],照射时温度为(4±2)℃;照射后标本通过细菌培养,透射电镜影像、核酸检测(LacZ基因片段)等方法观察核黄素光化学作用灭活大肠杆菌的效果。结果400—500nm可见光激发的核黄素光化学作用可以有效灭活大肠杆菌达6log;透射电镜影像示经核黄素光化学处理的大肠杆菌其拟核所在区域出现多个空泡;PCR显示经核黄素光化学处理的大肠杆菌其LacZ基因片段的复制被完全阻止。结论可见光激发的核黄素光化学作用可以有效灭活大肠杆菌,核酸是核黄素光化学产生病原体灭活作用的主要靶点。

The Research of riboflavin photochemical treatment with Escherichia coli

Objective In this research, pathogen inactivation effect and its mechanisms of riboflavin photochemical treatment with Escherichia coli is studied. Methods The 4.95 mL mixture of Escherichia coli and riboflavin were added into 5mL medical PVC transparent bags. Control groups were not exposed to light in the absence of riboflavin. Experimental groups were exposed to light (400-500 nm) in the present of riboflavin (100 umol/L) at a final dose of (8-32)J/cm~2. Lighting group were exposed to light (400-500 nm) in the absence of riboflavin. The effect and the mechanism of Escherichia coli inactivation was analyzed by bacteria cuhurvation, transmission electron technology and nucleic acid tests. Resuits 6 log of Escherichia coli was inactivated by rivoflavin photochemical treatment. Transmission electron photology showed that vacuoles were discovered in the nuclear area of riboflavin treated Escherichia coli. And the bacteria nucleic acid couldn't be amplified by PCR. Conclusion Riboflavin photochemical treatment with visible light can inactivate gscherichia coli and damage their nuclear acid. The research suggested Pathogen inactivation technology may be an efficient and feasible method in pathogens prevention in one step and supplied experimental supports for application in transfusion.