| Plasma is the raw material of blood products, and is also widely used in clinical work. If it had been contaminated by pathogen, a serious troubles would be happened. The safety of plasma have become one focus of medicine field. Although researchers have paid much attention to the checking and filtrating about the donor, For the reasons of mistaking supervisory management, limited sensitivity of detecting techniques, phenomenon of windows for virus presence and new pathogens are discovered between whiles, the problem of blood products contamination still can not be resolved effectively.Sterilization has been considered as the best measure to ensure a high level of safety applicable to plasma and its products. Since it contains a lot of proteins, the inactivation of virus in plasma is quite difficult. the effective approach has not been found yet. In this study, the phage f2 have been used as test virus. Through two parts of experiments: The inactivation of phage f2 in plasma by AO and UVA irradiation and the influence on major constituents of disinfected plasma. We explored the possibility of cooperation effect of photochemical technique produced by acridine orange (AO) and long-wave ultraviolet rays (UVA). The main results are described as follows:1. When the plasma was treated with AO alone, at the doses of 6μg/ml,7μg/ml and 10μg/ml for 5～30 min, the titers of bacteriophage f2 in plasma could reduced respectively by 3.27～4.53, 4.99～5.75, 6.11～7.53log.2. When the plasma was irradiated with UVA alone, the irradiation of UVA (4500J/m2-27000J/m2) could reduced the bacteriophage f2 in plasma by 1.21～2.63log. 3. The orthogonal assays indicated that, at the designed factors and levels, the AO doses was the main factor which influence the efficacy of inactivation of bacteriophage f2 in plasma. The irradiation time of UVA, the intensity of UVA and the doses of Rutin were not the main factors . 4. When be treated with AO and then irradiation by UVA, the titers of bacteriophage f2 in plasma were significantly reduced. The value of T/E of these two agents was higher than 1. This data indicated that UVA and AO have synergetic effects. After treated with 6μg/ml AO for 30mins than irradiation by 4500J/m2 UVA , the titers of bacteriophage f2 in plasma could reduced above 6log. 5. Observation by transmission electromicroscope showed that in plasma treated with AO and UVA at dosages effective for inactivating virus, most of the particle changed its shape, from spheriform become to spindle. The outlines of the virus were not clear and the electronic-density decreased. 6. Through examining by automatic biochemical analysis, SDS-PAGE electrophoresis, cross-immunoelectrophoresis, Enzyme-linked immunoadsordent assay, we found that after added the rutin, there was not significant change among the disinfected plasma, its contents of proteins and most biochemical parameters all remained within the normal ranges.Conclusion and prospect:1. After protection by 0.35mmol/L rutin, the cooperation treatment of 6 μg/ml AO and 4500J/m2 UVA irradiation could effectively inactivate the bacteriophage f2 in plasma and had little influence on the component of plasma. So we considered that the synergetic effects of AO and UVA is a utility methodbe used to inactivate the virus in plasma.2. With the advantages of low cost, simple in use, AO and UVA have great possibility for application to plasma disinfection. However, related research is just begining, and a lot of questions remain unresolved. Such as what is the regularity of inactivating in different viruses in plasma by AO and UVA ? Is the disinfected plasma poisonous to human body ? The systematic study on the influence of these factors on plasma components also needs to be carried out.
|
PDF Full Text Request