Laboratoire de Chimie Physique et Microbiologie pour l'Environnement (LCPME), UMR 7564 CNRS, Nancy-Université, 405 rue de Vandoeuvre, 54600 Villers-lès-Nancy, France.
J Virol Methods. 2012 Apr;181(1):18-24. doi: 10.1016/j.jviromet.2012.01.005. Epub 2012 Jan 30.
This study aimed to provide a tool for selecting the best approach to virological testing of bottled waters. Different methods were investigated. Method A examined the recovery of virus RNA following in situ lysis of virus particles in the aqueous phase and of those adhered to the bottle wall, method B examined the recovery of virus RNA following lysis of virus particles in the aqueous phase, and method C examined the recovery of intact virus particles. Method C generated the lowest genome recovery rate regardless of the water and virus type used, therefore comparison was mainly conducted between methods A and B.The effects of independent variables on the viral RNA recovery rate were determined by full factorial design. These independent variables included three waters (differing in mineral composition), four viruses (poliovirus 1, hepatitis A virus, Norovirus, and the MS2 phage), three incubation times (0, 10, and 20 days), and two methods (A and B). According to the results, each factor influenced the recovery rate of viral RNA with the exception of incubation time. Statistical analysis identified interactions between the factors. The strongest interactions involved the water and virus types, as well as the methods. The results suggested that method A should be used for the concentration and detection of hepatitis A virus, regardless of the divalent cation concentration of the bottled water. Method A was most suitable for water with the highest mineral content (divalent cation concentration of 250 mgL(-1)) and for the analysis of viruses capable of adsorbing onto the bottle walls (Poliovirus 1). Method B could be recommended for the analysis of water whose cation concentration is unknown.
本研究旨在提供一种用于选择最佳瓶装水病毒学检测方法的工具。考察了不同的方法。方法 A 考察了病毒颗粒在水相中和附着在瓶壁上的病毒颗粒原位裂解后病毒 RNA 的回收情况,方法 B 考察了病毒颗粒在水相中裂解后病毒 RNA 的回收情况,方法 C 考察了完整病毒颗粒的回收情况。无论使用何种水和病毒类型,方法 C 产生的基因组回收率最低,因此主要比较了方法 A 和方法 B。通过完全析因设计确定了独立变量对病毒 RNA 回收率的影响。这些独立变量包括三种水(矿物质成分不同)、四种病毒(脊髓灰质炎病毒 1、甲型肝炎病毒、诺如病毒和 MS2 噬菌体)、三种孵育时间(0、10 和 20 天)和两种方法(A 和 B)。结果表明,除了孵育时间外,每个因素都影响病毒 RNA 的回收率。统计分析确定了因素之间的相互作用。最强的相互作用涉及水和病毒类型以及方法。结果表明,无论瓶装水中二价阳离子浓度如何,都应使用方法 A 对甲型肝炎病毒进行浓缩和检测。方法 A 最适合矿物质含量最高的水(二价阳离子浓度为 250mgL(-1)) 和分析能够吸附在瓶壁上的病毒(脊髓灰质炎病毒 1)。对于阳离子浓度未知的水,可以推荐使用方法 B。