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Glass Wool Concentration Optimization for the Detection of Enveloped and Non-enveloped Waterborne Viruses.
Food and Environmental Virology ( IF 3.4 ) Pub Date : 2019-03-21 , DOI: 10.1007/s12560-019-09378-0
Albert Blanco 1, 2 , Islem Abid 3 , Nawal Al-Otaibi 3 , Francisco José Pérez-Rodríguez 1, 2 , Cristina Fuentes 1, 2 , Susana Guix 1, 2 , Rosa M Pintó 1, 2 , Albert Bosch 1, 2
Affiliation  

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.

中文翻译:

用于检测包膜和非包膜水性病毒的玻璃棉浓度优化。

一种非常经济实惠的病毒浓缩方法,该方法基于对玻璃棉的吸附洗脱和随后通过聚乙二醇6000(PEG)沉淀的再浓缩进行了优化,不仅可以回收非包膜病毒,而且可以回收包膜病毒。分别使用甲型肝炎病毒(HAV)和可传播的胃肠炎病毒(TGEV)替代裸露病毒和包膜病毒,以建立该方法。在小批量样品中的初步实验表明,两种类型的颗粒都易于吸附到带正电的玻璃棉上,但是通过标准洗脱(0.05M甘氨酸和3%的牛肉提取缓冲液,pH 9.5,洗脱效率极低),洗脱效率为7.2 HAV和TGEV分别为%和2.6%。为了提高包膜病毒的回收率,分析了洗脱液的几种变化:增加洗脱液的pH值,延长玻璃棉和洗脱液的接触时间,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下循环20分钟是选择的洗脱程序,在50 L水中,HAV和TGEV的效率分别为25.7%和18.8%。此外,采用20%PEG(而不是10%)进行病毒浓缩时,回收率分别提高了47%和51%。该优化程序用于检测利雅得Wadi Hanifa地表水中天然存在的HAV和冠状病毒。在38%的样本中检测到HAV,而一个样本的α冠状病毒呈阳性。
更新日期:2019-03-21
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