Assessment of the Applicability of Capsid-Integrity Assays for Detecting Infectious Norovirus Inactivated by Heat or UV Irradiation.
Assessment of the Applicability of Capsid-Integrity Assays for Detecting Infectious Norovirus Inactivated by Heat or UV Irradiation.
复制标题
衣壳完整性测定法检测热或紫外线照射灭活的传染性诺如病毒的适用性评估。
DOI：
10.1007/s12560-019-09390-4
复制
发表时间：
2019
影响因子：
3.4
通讯作者：
Walker DI
中科院分区：
文献类型：
--
作者：
Walker DI
人类诺如病毒是病毒性胃肠炎的主要原因。在缺乏用于常规分析感染性诺如病毒的实用培养技术的情况下，已经开发了几种方法，通过在RT-qPCR分析之前去除非活病毒来区分感染性和非感染性病毒。在这项研究中，两个这样的方法（RNA酶和猪胃粘蛋白），旨在消除病毒与受损衣壳（因此假定为非活的），评估其定量的能力，活的F-特异性RNA噬菌体（FRNAP）和人类诺如病毒灭活后，通过UV-C或热。结果发现，虽然两种方法都可以去除一部分非存活病毒，但仍有很大一部分非存活病毒有待通过RT-qPCR检测，导致高估了存活病毒群。然后开发了一种模型，以确定必须通过此类方法去除的非存活病毒的RT-qPCR可检测RNA的比例，以将高估降低至可接受的水平。在大多数情况下，必须去除几乎所有非存活病毒，以将存活率的对数高估降低到可接受的水平（例如低于0.5 log 10）。当开发替代预处理方法时，可以应用该模型，以确定它们的性能如何，以便与已建立的感染性检测方法相当。
Human noroviruses are the leading cause of viral gastroenteritis. In the absence of a practical culture technique for routine analysis of infectious noroviruses, several methods have been developed to discriminate between infectious and non-infectious viruses by removing non-viable viruses prior to analysis by RT-qPCR. In this study, two such methods (RNase and porcine gastric mucin) which were designed to remove viruses with compromised capsids (and therefore assumed to be non-viable), were assessed for their ability to quantify viable F-specific RNA bacteriophage (FRNAP) and human norovirus following inactivation by UV-C or heat. It was found that while both methods could remove a proportion of non-viable viruses, a large proportion of non-viable virus remained to be detected by RT-qPCR, leading to overestimations of the viable population. A model was then developed to determine the proportion of RT-qPCR detectable RNA from non-viable viruses that must be removed by such methods to reduce overestimation to acceptable levels. In most cases, nearly all non-viable virus must be removed to reduce the log overestimation of viability to within levels that might be considered acceptable (e.g. below 0.5 log10). This model could be applied when developing alternative pre-treatment methods to determine how well they should perform to be comparable to established infectivity assays.