Anderson-Coughlin Brienna L, Kniel Kalmia E
Department of Animal and Food Sciences, University of Delaware, Newark, DE 19716, USA.
Methods Protoc. 2019 Jun 30;2(3):55. doi: 10.3390/mps2030055.
The variability of environmental water samples impacts the allowance of one method to be universally ideal for all water types and volumes. Surface and reclaimed waters can be used for crop irrigation and may be referred to as non-traditional irrigation waters as these water types may be associated with a higher risk of microbial contamination compared to groundwater. These waters are typically more microbially and chemically complex than groundwater and have a higher risk of viral contamination. To detect viruses in these water types, an infinite number of variations can be made to traditional recovery methods. This protocol was developed based on a commonly used virus adsorption and elution (VIRADEL) method. Additional steps were included to simplify and efficiently reduce particulates in the viral concentrate and remove DNA from eluted nucleic acids prior to detection. Method alterations allow for volumes up to 40 liters to be processed with consistent recovery of enteric viruses including Aichi virus, hepatitis A virus, and noroviruses belonging to genogroups GI and GII. No inhibition was observed among either surface or reclaimed water samples. This protocol could be utilized in the monitoring of a wide array of irrigation water sources throughout irrigation processes.
环境水样的变异性影响了一种方法对所有水类型和水量都普遍理想的可能性。地表水和再生水可用于作物灌溉,并且可能被称为非传统灌溉用水,因为与地下水相比,这些水类型可能与更高的微生物污染风险相关。这些水通常在微生物和化学方面比地下水更复杂,并且有更高的病毒污染风险。为了检测这些水类型中的病毒,可以对传统的回收方法进行无数种变化。本方案是基于一种常用的病毒吸附和洗脱(VIRADEL)方法开发的。还包括了额外的步骤,以简化并有效减少病毒浓缩物中的颗粒,并在检测前从洗脱的核酸中去除DNA。方法的改变允许处理高达40升的水样,并能一致地回收包括爱知病毒、甲型肝炎病毒以及属于基因群GI和GII的诺如病毒在内的肠道病毒。在地表水或再生水样本中均未观察到抑制作用。本方案可用于在整个灌溉过程中监测各种灌溉水源。