陡脉冲电场选择性灭活血液中病原体效果观察

Effect of steep pulsed electric field on pathogen inactivation in blood

目的：观察纳秒级陡脉冲在10 ns脉宽参数下对病原体的杀伤效应,以及对红细胞的损伤作用,研究高压脉冲电场应用于血液病原体灭活的可能性。方法：以大肠埃细菌为研究对象,采用脉冲参数40～80 k V/cm、脉宽10 ns、重复频率1 Hz的陡脉冲进行实验处理,实验菌株活力以MTT法进行检测,三波长法检测处理前后的血液样品的游离血红蛋白（FHb）浓度。结果：在电场强度40 k V/cm、10个脉冲作用下活性抑制率达到54.5%,不同介质中脉冲数对细菌活性均有显著影响（P〈0.05）;不同脉冲电场强度越大其抑制率有所升高,但均无统计学意义（P〉0.05）;无菌条件下,脉冲数对红细胞的影响显著（P〈0.05）;有菌条件下,脉冲数及电场强度对溶血仍有影响,但统计结果不显著（电场强度P=0.159,脉冲数P=0.278）;FHb在10个脉冲数的作用下,没有显著增加,甚至比对照组更低。结论：纳秒级陡脉冲电场具备较强的杀菌能力,且在一定参数条件下不对血细胞产生明显的损伤作用,具备选择性杀伤的潜在作用。

Objective To research on the possibility of applying high voltage pulsed electric field for pathogen inactivation in blood by observing the effects of steep pulsed electric field （SPEF） at 10 ns pulse width on pathogen inactivation, and studying the adverse influence on the red blood cell. Methods The Escherichia coli was treated by the steep pulse with 40- 80 kV/cm, 10 ns pulse width, repetition frequency of 1 Hz. The bacterial activity was detected by MTT method, and the concentration of free hemoglobin （FHb） in blood sample was detected before and after three wavelength spectrophotometry. Results The inhibition ratio of bacterial activity was 54.5% with the treatment of 10 pulses at electric field intensity of 40 kV/cm. In different media, the number of pulses showed significant effects on the bacterial activity （P〈0.05）. The inhibited ratio of bacterial activity increased with the increasing electric filed density, without any statistical significance （P〉0.05）. Under the condition without bacteria, the number of pulses showed significant impacts on red blood cells （P〈0.05）; under the condition with bacteria, the number of pulses and electric filed density had effects on red blood cells, without significant statistical results （P=0.278, P=-0.159）. With the treatment of 10 pluses, the FHb concentration did not significantly increased, even lower than the FHb concentration of control group. Conclusion The nanosecond SPEF has strong ability of sterilization, without causing significant adverse effects on red blood cells at certain parameter conditions, which indicates a potential application of SPEF in selectively inactivating pathogens in blood.