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使用磁性硅胶珠从冷冻树莓中提取诺如病毒。

Norovirus Extraction from Frozen Raspberries Using Magnetic Silica Beads.

机构信息

Canadian Food Inspection Laboratory (CFIA), St. Hyacinthe Laboratory, Food Virology, Saint Hyacinthe, QC, Canada.

Agriculture and Agri-Food Canada (AAFC), St. Hyacinthe Research and Development Centre, Saint Hyacinthe, QC, Canada.

出版信息

Food Environ Virol. 2021 Jun;13(2):248-258. doi: 10.1007/s12560-021-09466-0. Epub 2021 Mar 2.

Abstract

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.

摘要

人诺如病毒(HuNoV)是全球急性胃肠炎的主要病因之一。冷冻覆盆子已与几起与 HuNoV 相关的食源性暴发事件有关。然而,从冷冻覆盆子中提取 HuNoV RNA 仍然具有挑战性。回收产量低,实时定量逆转录聚合酶链反应(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硅胶珠的新方法,用于提取冷冻覆盆子中添加的 HuNoV。用磁硅胶珠和参考 ISO 15216-1:2017 方法均观察到相对较低的回收产量。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。减少 RT-qPCR 抑制对于减少不确定 HuNoV 检测数量很重要,从而增加评估冷冻覆盆子中 HuNoV 流行率的能力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ac14/8116234/8badc4665885/12560_2021_9466_Fig1_HTML.jpg

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