Laboratory of Comparative and Environmental Virology, Oswaldo Cruz Institute, Fiocruz, Rio de Janeiro-RJ, Brazil.
Laboratory of Comparative and Environmental Virology, Oswaldo Cruz Institute, Fiocruz, Rio de Janeiro-RJ, Brazil.
J Virol Methods. 2018 Oct;260:49-55. doi: 10.1016/j.jviromet.2018.07.001. Epub 2018 Jul 4.
This study aimed to assess viral elution-concentration methods for recovering noroviruses from deli meats. Spiking experiments were conducted to evaluate the recovery success rates and recovery efficiencies of human norovirus (NoV) GI and GII and murine norovirus 1 (MNV-1) using polyethylene glycol (PEG 6000) precipitation, skimmed milk flocculation (SMF), TRIzol reagent, and a combination of PEG/TRIzol and SMF/TRIzol methods. Molecular analysis using reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) revealed TRIzol as the best method to be used for viral recovery from ham with medium recovery rates of 37.6% for NoV GI and 50.1% for NoV GII. Viral recovery from turkey meat showed medium recovery rates of 14.4% for NoV GI and 8.9% for NoV GII. For MNV-1, the rates varied from 0.5% to 80.8% not only according to the matrix but also with the associated virus and its inoculum (NoV GI or GII). The monitoring of commercial samples obtained in the Great Metropolitan region of Rio de Janeiro in order to demonstrate the occurrence of NoV GI and GII contamination in both matrices was also performed in 60 samples. NoV GI or GII were not detected in any samples, while MNV-1 used as the sample process control viruswas successfully recovered in 100% of samples.
本研究旨在评估从熟食肉类中回收诺如病毒的病毒洗脱浓缩方法。通过聚乙二醇（PEG 6000）沉淀、脱脂乳絮凝（SMF）、TRIzol 试剂以及 PEG/TRIzol 和 SMF/TRIzol 联合方法，进行了刺血实验以评估人类诺如病毒（NoV）GI 和 GII 以及鼠诺如病毒 1（MNV-1）的回收成功率和回收率。使用逆转录定量聚合酶链反应（RT-qPCR）进行分子分析显示，TRIzol 是从火腿中回收病毒的最佳方法，NoV GI 的回收率中等，为 37.6%，NoV GII 的回收率为 50.1%。从火鸡肉中回收病毒的回收率中等，NoV GI 的回收率为 14.4%，NoV GII 的回收率为 8.9%。对于 MNV-1，回收率不仅取决于基质，还取决于相关病毒及其接种物（NoV GI 或 GII），从 0.5%到 80.8%不等。还对从里约热内卢大都市区获得的商业样本进行了监测，以证明这两种基质中都存在 NoV GI 和 GII 污染。在 60 个样本中均未检测到 NoV GI 或 GII，而用作样品处理对照病毒的 MNV-1 在 100%的样本中均被成功回收。
