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Norovirus Extraction from Frozen Raspberries Using Magnetic Silica Beads
Food and Environmental Virology ( IF 4.1 ) Pub Date : 2021-03-02 , DOI: 10.1007/s12560-021-09466-0
Philippe Raymond 1 , Sylvianne Paul 1 , André Perron 1 , Louise Deschênes 2
Affiliation  

Human noroviruses (HuNoV) are among the main causes of acute gastroenteritis worldwide. Frozen raspberries have been linked to several HuNoV food-related outbreaks. However, the extraction of HuNoV RNA from frozen raspberries remains challenging. Recovery yields are low, and real-time quantitative reverse transcriptase PCR (RT-qPCR) inhibitors limit the sensitivity of the detection methodologies. A new approach using fine magnetic silica beads was developed for the extraction of HuNoV spiked on frozen raspberries. Relatively low recovery yields were observed with both the magnetic silica bead and the reference ISO 15216-1:2017 methods. High RT-qPCR inhibition was observed with the ISO 15216-1:2017 recommended amplification kit but could be reduced by using an alternative kit. Reducing RT-qPCR inhibition is important to limit the number of inconclusive HuNoV assays thus increasing the capacity to assess the HuNoV prevalence in frozen raspberries.



中文翻译:

使用磁性二氧化硅珠从冷冻覆盆子中提取诺如病毒

人类诺如病毒 (HuNoV) 是全球急性胃肠炎的主要原因之一。冷冻覆盆子与几起与 HuNoV 食品相关的疫情有关。然而,从冷冻覆盆子中提取 HuNoV RNA 仍然具有挑战性。回收率低,实时定量逆转录酶 PCR (RT-qPCR) 抑制剂限制了检测方法的灵敏度。开发了一种使用细磁性二氧化硅珠的新方法,用于提取添加在冷冻覆盆子上的 HuNoV。使用磁性硅胶珠和参考 ISO 15216-1:2017 方法均观察到相对较低的回收率。使用 ISO 15216-1:2017 推荐的扩增试剂盒观察到高 RT-qPCR 抑制,但可以通过使用替代试剂盒来降低。

更新日期:2021-03-02
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