Fundação Oswaldo Cruz-Fiocruz, Instituto Oswaldo Cruz, Laboratório de Morfologia e Morfogênese Viral, Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz-Fiocruz, Instituto Oswaldo Cruz, Laboratório de Patologia, Rio de Janeiro, RJ, Brasil.
Mem Inst Oswaldo Cruz. 2023 Aug 28;118:e230090. doi: 10.1590/0074-02760230090. eCollection 2023.
According to the last 2023 Monkeypox (Mpox) Outbreak Global Map from the Centres for Disease Control and Prevention (CDC), more than 100 countries with no Mpox infection report cases. Brazil stands out in this group and is the second country with the highest number of cases in the last outbreak.
To contribute to knowledge of the virus infection effects in a cellular model, which is important for diagnosis infections not yet included in a provider´s differential diagnosis and for developing viral inhibition strategies.
We describe a virus isolation protocol for a human clinical sample from a patient from Brazil, the viral growth in a cell model through plaque forming units (PFU) assay, reverse transcriptase polymerase chain reaction (RT-PCR) and transmission electron microscopy (TEM).
We follow the viral isolation in Vero cell culture from a Mpox positive clinically diagnosed sample and show the infection effects on cellular structures using a TEM.
Understanding the impact of viral growth on cellular structures and its replication kinetics may offer better strategies for the development of new drugs with antiviral properties.
根据疾病控制与预防中心(CDC)发布的 2023 年猴痘(Mpox)疫情全球地图,已有超过 100 个无猴痘感染报告的国家。巴西在这些国家中表现突出,是上一次疫情中报告病例数第二多的国家。
为病毒感染在细胞模型中的影响提供知识,这对于诊断尚未纳入提供者鉴别诊断的感染以及开发病毒抑制策略非常重要。
我们描述了一种从巴西患者的人类临床样本中进行病毒分离的方案,通过噬斑形成单位(PFU)测定、逆转录聚合酶链反应(RT-PCR)和透射电子显微镜(TEM)来检测病毒在细胞模型中的生长情况。
我们遵循从猴痘阳性临床诊断样本中在 Vero 细胞培养物中的病毒分离,并使用 TEM 显示感染对细胞结构的影响。
了解病毒生长对细胞结构的影响及其复制动力学,可能为开发具有抗病毒特性的新药提供更好的策略。