当前位置: X-MOL 学术Food Environ. Virol. › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
Comparison of Extraction Methods for the Detection of Tick-Borne Encephalitis Virus RNA in Goat Raw Milk and Cream Cheese
Food and Environmental Virology ( IF 4.1 ) Pub Date : 2022-09-20 , DOI: 10.1007/s12560-022-09535-y
Irene Müller 1 , Nadine Althof 1 , Bernd Hoffmann 2 , Christine Klaus 3 , Katja Schilling-Loeffler 1 , Alexander Falkenhagen 1 , Reimar Johne 1
Affiliation  

Infection with the tick-borne encephalitis virus (TBEV) can cause meningitis, meningoencephalitis and myelitis in humans. TBEV is an enveloped RNA virus of the family Flaviviridae, which is mostly transmitted via tick bites. However, transmission by consumption of virus-contaminated goat raw milk and goat raw milk products has also been described. Only a few methods have been reported for the detection of TBEV in food so far. Here, we compare different virus extraction methods for goat raw milk and goat raw milk cream cheese and subsequent detection of TBEV-RNA by RT-qPCR. Langat virus (LGTV), a naturally attenuated TBEV strain, was used for artificial contamination experiments. Mengovirus and the human coronavirus 229E were compared to assess their suitability to serve as internal process controls. Out of three tested extraction protocols for raw milk, sample centrifugation followed by direct RNA extraction from the aqueous interphase yielded the best results, with a recovery rate (RR) of 31.8 ± 4.9% for LGTV and a detection limit of 6.7 × 103 LGTV genome copies/ml. Out of two methods for cream cheese, treatment of the samples with TRI Reagent® and chloroform prior to RNA extraction showed the best RR of 4.7 ± 1.6% for LGTV and a detection limit of 9.4 × 104 LGTV genome copies/g. RRs of Mengovirus and LGTV were similar for both methods; therefore, Mengovirus is suggested as internal process control virus. The developed methods may be useful for screening or surveillance studies, as well as in outbreak investigations.



中文翻译:

山羊原料奶和奶油干酪中蜱传脑炎病毒 RNA 提取方法的比较

蜱传脑炎病毒 (TBEV) 感染可引起人类脑膜炎、脑膜脑炎和脊髓炎。TBEV 是黄病毒科的一种包膜 RNA 病毒,主要通过蜱虫叮咬传播。然而,也有通过食用受病毒污染的山羊原料奶和山羊原料奶制品传播的报道。迄今为止,检测食品中结核病病毒的方法很少见。在这里,我们比较了山羊原奶和山羊原奶奶油奶酪的不同病毒提取方法以及随后通过 RT-qPCR 检测 TBEV-RNA。Langat 病毒 (LGTV) 是一种自然减毒的 TBEV 菌株,用于人工污染实验。对 Mengovirus 和人类冠状病毒 229E 进行了比较,以评估它们作为内部过程控制的适用性。在三种经过测试的原料奶提取方案中,样品离心然后从水相间直接提取 RNA 产生了最好的结果,回收率 (RR) 为 31.8 ± 4。3 LGTV 基因组拷贝/ml。在用于奶油干酪的两种方法中,在提取 RNA 之前用 TRI Reagent® 和氯仿处理样品显示 LGTV 的最佳 RR 为 4.7 ± 1.6%,检测限为 9.4 × 10 4 LGTV 基因组拷贝/ g。两种方法的 Mengovirus 和 LGTV 的 RR 相似;因此,建议将 Mengovirus 作为内部过程控制病毒。开发的方法可能对筛选或监测研究以及暴发调查有用。

更新日期:2022-09-21
down
wechat
bug