目的:开发了连续骤冷流(CQF)反应器,以在少于一秒的反应时间内收集样品。该反应器用于确定脊髓灰质炎病毒的臭氧消毒动力学,并研究EMA-qPCR是否可以评估臭氧消毒后的病毒感染性。
方法:在开发的CQF中进行脊髓灰质炎病毒的臭氧消毒,在0·8和0·25mgl-1的臭氧浓度下,在0·7-5·0s范围内测试了消毒动力学。失活,对病毒基因组的损伤和对衣壳完整性的损伤通过噬斑测定来确定,定量逆转录聚合酶链反应(RT-qPCR)和单叠氮乙锭处理与RT-qPCR(EMA-qPCR),分别。
结果:通过使用CQF,在反应时间为0·7s,臭氧浓度为0·08和0·25mgl-1时,观察到2·18和2·76log10的减少,分别,其次是拖尾。效率因子Hom模型比Chick-Watson模型对脊髓灰质炎病毒1型的臭氧消毒动力学拟合更好,或修改后的Chick-Watson模型。在<2·0s的反应时间和0·08和0·25mgl-1的臭氧浓度下,在RT-qPCR和EMA-qPCR测定之间观察到的动力学相似。在反应时间>5s时,与稳定的RT-qPCR结果相比,通过EMA-qPCR评估的病毒浓度降低.两种检测方法仍然低估了病毒的灭活作用。
结论:简单开发的反应器可用于研究病毒臭氧消毒动力学,并在非常短的暴露时间内阐明灭活特性或机制。
结论:开发的CQF反应器有利于更好地理解臭氧灭活病毒,该反应器可以更好地阐明消毒动力学和机理,为今后的研究提供依据。这项工作对EMA/PMA-qPCR在臭氧消毒后评估病毒感染性的应用和局限性的现有知识做出了重要贡献。
OBJECTIVE: A continuous quench-flow (CQF) reactor was developed to collect samples at the reaction times of less than one second. The reactor is applied to determine ozone disinfection kinetics of poliovirus and to study whether EMA-qPCR can assess the viral infectivity after ozone disinfection.
METHODS: Ozone disinfection of poliovirus was conducted in the developed CQF, and the disinfection kinetics were tested in the range of 0·7-5·0 s at ozone concentration of 0·08 and 0·25 mg l-1 . Inactivation, damage on viral genome and damage on capsid integrity were determined by plaque assay, quantitative reverse transcription polymerase chain reaction (RT-qPCR) and ethidium monoazide treatment coupled with RT-qPCR (EMA-qPCR), respectively.
RESULTS: By using CQF, 2·18 and 2·76 log10 reductions were observed at the reaction time of 0·7 s and ozone concentration of 0·08 and 0·25 mg l-1 , respectively, followed by tailing. Ozone disinfection kinetics of poliovirus 1 were better fit by the efficiency factor Hom model than by the Chick-Watson model, or the modified Chick-Watson model. Kinetics observed were similar between RT-qPCR and EMA-qPCR assays at the reaction times of <2·0 s and ozone concentrations of 0·08 and 0·25 mg l-1 . At reaction times > 5 s, viral concentration evaluated by EMA-qPCR was reduced in comparison to stable RT-qPCR results. Both assays still underestimated the virus inactivation.
CONCLUSIONS: The simple developed reactor can be used to investigate viral ozone disinfection kinetics and to elucidate inactivation characteristics or mechanisms at very short exposure times.
CONCLUSIONS: The developed CQF reactor is beneficial for better understanding of virus inactivation by ozone, and the reactor can be used to better elucidate disinfection kinetics and mechanisms for future research. This work constitutes an important contribution to the existing knowledge of the application and limitation of the EMA/PMA-qPCR to assess virus infectivity after ozone disinfection.