Glass Wool Concentration Optimization for the Detection of Enveloped and Non-enveloped Waterborne Viruses
Glass Wool Concentration Optimization for the Detection of Enveloped and Non-enveloped Waterborne Viruses
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DOI：
10.1007/s12560-019-09378-0
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发表时间：
2019-06-01
影响因子：
3.4
通讯作者：
Bosch, Albert
中科院分区：
文献类型：
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作者：
Blanco, Albert;Abid, Islem;Bosch, Albert
优化了一种非常经济实惠的病毒浓缩方法，该方法基于吸附-洗脱至玻璃棉，随后通过聚乙二醇6000（PEG）沉淀再浓缩，不仅可回收无包膜病毒，还可回收包膜病毒。甲型肝炎病毒（HAV）和传染性胃肠炎病毒（TGEV）分别作为裸病毒和包膜病毒的替代物，以建立方法学。在小体积样品中的初始实验表明，这两种类型的颗粒容易吸附到带正电荷的玻璃棉，但通过标准洗脱与0.05 M甘氨酸与3%的牛肉提取物缓冲液，pH 9.5，洗脱效率为7.2%和2.6%，分别为HAV和TGEV，从它差分离。为了提高包膜病毒的回收率，测定了洗脱中的几种修改：提高洗脱pH值，延长玻璃棉和洗脱液接触时间，添加去污剂，或通过再循环或搅拌进行洗脱。考虑到实用性和性能，洗脱液在pH 11.0下再循环20 min是选择的洗脱程序，在50 L水中对HAV和TGEV的效率分别为25.7%和18.8%。此外，使用20% PEG代替10%用于病毒再浓缩，将回收率分别提高至47%和51%。优化的程序被应用于检测自然发生的甲型肝炎病毒和冠状病毒在地表水的瓦迪哈尼法，利雅得。在38%的样本中检测到HAV，而一个样本对冠状病毒呈阳性。这种廉价的病毒检测系统能够全面监测水样中存在的病毒。
An extremely affordable virus concentration method based on adsorption-elution to glass wool and subsequent reconcentration through polyethylene glycol 6000 (PEG) precipitation was optimized to recover not only non-enveloped viruses but also enveloped viruses. Hepatitis A virus (HAV) and transmissible gastroenteritis virus (TGEV) were employed as surrogates for naked and enveloped viruses, respectively, to set up the methodology. Initial experimentation in small-volume samples showed that both types of particles readily adsorbed to the positively charged glass wool but were poorly detached from it through standard elution with 0.05 M glycine with 3% of beef extract buffer, pH 9.5, with elution efficiencies of 7.2% and 2.6%, for HAV and TGEV, respectively. To improve the recovery of enveloped viruses, several modifications in the elution were assayed: increasing the elution pH, extending glass wool and eluent contact time, adding a detergent, or performing the elution by recirculation or under agitation. Considering practicability and performance, recircularization of the eluent at pH 11.0 for 20 min was the elution procedure of choice, with efficiencies of 25.7% and 18.8% for HAV and TGEV in 50 L of water. Additionally, employing 20% PEG instead of 10% for virus reconcentration improved recoveries up to 47% and 51%, respectively. The optimized procedure was applied to detect naturally occurring HAV and coronaviruses in surface water of Wadi Hanifa, Riyadh. HAV was detected in 38% of the samples, while one sample was positive for an alphacoronavirus. This cheap virus detection system enables the comprehensive surveillance of viruses present in water samples.