Wang J, Mauser A, Chao S-F, Remington K, Treckmann R, Kaiser K, Pifat D, Hotta J
Bayer Healthcare, Research Triangle Park, NC, USA.
Vox Sang. 2004 May;86(4):230-8. doi: 10.1111/j.0042-9007.2004.00485.x.
Ultraviolet-C (UVC) irradiation is a viral-inactivation method that was dismissed by many plasma fractionators as a result of the potential for protein damage and the difficulty in delivering uniform doses. A reactor with novel spiral flow hydraulic mixing was recently designed for uniform and controlled UVC treatment. The objective of this study was to investigate virus inactivation and protein recovery after treatment through the new reactor.
Virus- and mock-spiked Alpha1-proteinase inhibitor (Alpha1-PI) solutions were treated with UVC. The virus samples were assayed for residual infectivity and amplified by the polymerase chain reaction (PCR). The mock-spiked samples were assayed for protein integrity.
Greater than 4 log10 of all test viruses were inactivated, regardless of the type of nucleic acid or presence of an envelope. Unlike previous studies, viruses with the smallest genomes were found to be those most sensitive to UVC irradiation, and detection of PCR amplicons > or = 2.0 kb was correlated to viral infectivity. Doses that achieved significant virus inactivation yielded recovery of > 90% protein activity, even in the absence of quenchers.
The results demonstrate the effectiveness of UVC treatment, in the novel reactor, to inactivate viruses without causing significant protein damage, and confirm the utility of large PCR amplicons as markers for infectious virus.
紫外线C(UVC)照射是一种病毒灭活方法,但由于存在蛋白质损伤的可能性以及难以实现均匀剂量输送,许多血浆分馏器已不再使用该方法。最近设计了一种具有新型螺旋流液压混合功能的反应器,用于进行均匀且可控的UVC处理。本研究的目的是通过新反应器研究处理后的病毒灭活情况和蛋白质回收率。
用UVC处理添加了病毒和模拟病毒的α1-蛋白酶抑制剂(Alpha1-PI)溶液。对病毒样本检测残留感染力,并通过聚合酶链反应(PCR)进行扩增。对添加了模拟病毒的样本检测蛋白质完整性。
所有测试病毒的灭活程度均超过4个对数10,无论核酸类型或是否有包膜。与以往研究不同,发现基因组最小的病毒对UVC照射最为敏感,且PCR扩增子>或=2.0 kb的检测与病毒感染力相关。即使在没有淬灭剂的情况下,实现显著病毒灭活的剂量也能使蛋白质活性回收率>90%。
结果表明,在新型反应器中,UVC处理能有效灭活病毒且不会造成显著的蛋白质损伤,并证实了大PCR扩增子作为传染性病毒标志物的实用性。