Melgaço Fabiana Gil, Victoria Matias, Corrêa Adriana Abreu, Ganime Ana Carolina, Malta Fábio Correia, Brandão Marcelo Luiz Lima, de Mello Medeiros Valéria, de Oliveira Rosas Carla, Bricio Silvia Maria Lopes, Miagostovich Marize Pereira
Laboratory of Comparative and Environmental Virology, Oswaldo Cruz Institute, Fiocruz, Rio de Janeiro-RJ, Brazil.
Laboratory of Molecular Virology, Regional Norte, CENUR Noroeste, Universidad de la República, Salto, Uruguay.
Int J Food Microbiol. 2016 Jan 18;217:14-9. doi: 10.1016/j.ijfoodmicro.2015.10.005. Epub 2015 Oct 9.
Skimmed milk organic flocculation method was adapted, optimized and compared with polyethylene glycol (PEG) precipitation and filtration methods for recovering viruses from a strawberry matrix. Spiking experiments with norovirus genogroup II genotype 4 (NoV GII.4) and murine norovirus 1 (MNV-1) demonstrated that the organic flocculation method associated with a glycine elution buffer, filter bag and cetyltrimethylammonium bromide (CTAB) showed a recovery percentage of 2.5 and 32 times higher than PEG precipitation and filtration methodologies for NoV recovering. Furthermore, this method was used for investigating NoV and human adenoviruses (HAdVs) in 90 samples of fresh strawberries commercialized in Rio de Janeiro markets. NoV GI and GII were not detected in those samples and MNV-1, used as internal process control (IPC), was recovered in 95.5% (86) of them. HAdVs were detected in 18 (20.0%) samples and characterized by nucleotide sequencing as Human Mastadenovirus specie F and as type specie HAdV-2. Bacterial analysis did not detect Salmonella spp. and Listeria monocytogenes, however, 3.3% of fecal coliforms were detected in those samples. These results indicate the organic flocculation method as an alternative for recovering enteric viruses from strawberries, emphasizing a need for virus surveillance in food matrices.
采用脱脂牛奶有机絮凝法，并对其进行优化，同时与聚乙二醇（PEG）沉淀法和过滤法进行比较，以从草莓基质中回收病毒。用诺如病毒II基因组4型（NoV GII.4）和鼠诺如病毒1型（MNV-1）进行加标实验表明，与甘氨酸洗脱缓冲液、滤袋和十六烷基三甲基溴化铵（CTAB）相结合的有机絮凝法，在回收NoV时的回收率比PEG沉淀法和过滤法分别高2.5倍和32倍。此外，该方法用于调查里约热内卢市场上商业化的90份新鲜草莓样本中的NoV和人腺病毒（HAdV）。在这些样本中未检测到NoV GI和GII，用作内部过程对照（IPC）的MNV-1在95.5%（86份）的样本中被回收。在18份（20.0%）样本中检测到HAdV，并通过核苷酸测序鉴定为人 Mastadenovirus 种F和HAdV-2型种。细菌分析未检测到沙门氏菌属和单核细胞增生李斯特菌，然而，在这些样本中检测到3.3%的粪大肠菌群。这些结果表明有机絮凝法可作为从草莓中回收肠道病毒的一种替代方法，强调了对食品基质中病毒监测的必要性。
