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目的探讨噻唑橙(TO)光化学法对红细胞中病毒及细菌灭活效果。方法以伪狂犬病毒(PRV)和辛德毕斯(Sindbis)病毒为指示病毒,分别加入红细胞比积为20%的红细胞悬液中,TO孵育1 h后做476 nm光照射处理;研究病毒灭活动力学特征;做TO浓度(C)、光照强度(I)、光照时间(T)三因素三水平正交试验确定最优灭活条件;最优条件下检测裸照、密闭血袋中、密闭血袋充氧后的病毒灭活效果;最优条件下检测血液污染常见细菌小肠结肠炎耶尔森氏菌、金黄色葡萄球菌及表皮葡萄球菌灭活效果。结果病毒灭活动力学研究显示:在60μmol/LTO、1.06E-01 w·m-2·nm-1光强条件下,照射5 min可将大部分PRV和Sindbis病毒灭活,20 min灭活作用达峰值,分别为5.88和5.12 LogTCID50;正交试验显示:裸照时PRV及Sindbis病毒灭活最优条件均为I=1.33E-01 w·m-2·nm-1,T=20 min,C=80μmol/L;在此条件下,可灭活红细胞中PRV≥6.13LogTCID50(裸照,n=4)、(4.75±0.62)LogTCID50(密闭血袋,n=4)、(6.06±0.16)LogTCID50(密闭血袋充氧,n=4);可灭活红细胞中Sindbis病毒(5.41±0.12)LogTCID50(裸照,n=4)、(3.72±0.77)LogTCID50(密闭血袋,n=4)、(5.76±0.25)LogTCID50(密闭血袋充氧,n=4)。在此条件下细菌灭活效果:小肠结肠炎耶尔森氏菌(5.91±0.13)Log10、金黄色葡萄球菌>6.8Log10、表皮葡萄球菌>6.0 Log10。结论 TO光化学法可有效灭活红细胞中病毒,有氧情况下(裸照或充氧)病毒灭活效果好于密闭无氧。TO光化学法可有效灭活红细胞中细菌。
Objective To investigate the inactivation effect of thiazole orange (TO) photochemistry on virus and bacteria in erythrocytes. Methods The PRV and Sindbis viruses were used as the indicator viruses and were respectively added to the erythrocyte suspensions with 20% hematocrit. After incubated for 1 h at TO, they were irradiated with 476 nm light. The virus inactivation kinetics The optimum inactivation conditions were determined by three factors and three levels of orthogonal test including TO concentration (C), light intensity (I) and light time (T). In the optimal conditions, Oxygen inactivation effect after the virus; the best detection of blood contamination common bacteria Yersinia enterocolitica, Staphylococcus aureus and Staphylococcus epidermidis inactivation. Results The kinetics of virus inactivation showed that most of the PRV and Sindbis viruses were inactivated at 60 μmol / L and 1.06E-01 w · m-2 · nm-1 for 5 min and were inactivated by 20 min (5.88 and 5.12 LogTCID50 respectively). The orthogonal test showed that the optimum conditions of inactivation of PRV and Sindbis virus in nude were I = 1.33E-01 w · m-2 · nm-1, T = 20 min, C = 80μmol / L. Under these conditions, PRV≥6.13LogTCID50 (nude, n = 4), (4.75 ± 0.62) LogTCID50 (closed blood bag, n = 4), (6.06 ± 0.16) LogTCID50 (5.41 ± 0.12) LogTCID50 (nude, n = 4), (3.72 ± 0.77) LogTCID50 (confined blood bag, n = 4), ± 0.25) LogTCID50 (closed blood bag oxygenation, n = 4). Inactivation of bacteria under these conditions: Yersinia enterocolitica (5.91 ± 0.13) Log10, Staphylococcus aureus> 6.8Log10, Staphylococcus epidermidis> 6.0 Log10. Conclusion TO photochemical method can effectively inactivate the virus in erythrocytes, aerobic conditions (nude or oxygenated) virus inactivation better than closed anaerobic. TO photochemical method can effectively inactivate bacteria in red blood cells.