Chung Hee Chun, Kim Sung Jae, Hwang Su Jin, Park Sung Hoon, Park Kyoung Min, Chung Hyeon Woo, Ko Si Hwan, Park Dong Il, Shim Jun-Yeop, Nguyen Van Giap, Lee Jae Myun
Department of Microbiology and Immunology, Institute for Immunology and Immunological Diseases, Yonsei University College of Medicine, Seoul, Republic of Korea.
Department of Companion Animal Health, Kyungbok University, Namyangju, Republic of Korea.
Front Microbiol. 2025 Aug 26;16:1654224. doi: 10.3389/fmicb.2025.1654224. eCollection 2025.
Pediatric isolates of SARS-CoV-2 remain underrepresented in virologic studies, despite their importance for understanding viral diversity and therapeutic responses.
Nasal swab and saliva samples were collected from pediatric COVID-19 patients. Viral isolation was attempted in Vero cells through five blind passages. Replication was assessed by digital RT-PCR, while cytopathic effects were observed microscopically. Genomic sequencing was conducted using next-generation sequencing, and antiviral activity was evaluated for Remdesivir, Molnupiravir, and Nirmatrelvir.
A SARS-CoV-2 strain, designated N15, was successfully isolated from a pediatric nasal swab. The isolate replicated efficiently in Vero cells with kinetics comparable to B.1 and B.1.1.529 lineages. Cytopathic effects appeared within 48 h post-infection, marked by aggregates of dead cells. Genomic analysis classified N15 within lineage 19B, showing 99.9% similarity to the Wuhan-Hu-1 strain. Rare mutations were identified: N709S in the spike protein and T11M in the E protein. Antiviral testing revealed effective inhibition by Remdesivir, Molnupiravir, and Nirmatrelvir, with varying IC50 values across cell types.
The pediatric N15 isolate represents a unique 19B lineage virus, retaining ancestral genomic features while harboring rare mutations. Its efficient replication and drug sensitivity underscore its value as a reference strain for comparative studies against circulating variants and for evaluating antiviral efficacy.
尽管儿童感染的严重急性呼吸综合征冠状病毒2(SARS-CoV-2)毒株对于理解病毒多样性和治疗反应具有重要意义,但在病毒学研究中其代表性仍然不足。
从患有新型冠状病毒肺炎(COVID-19)的儿童患者中采集鼻拭子和唾液样本。尝试在Vero细胞中通过五次盲传进行病毒分离。通过数字逆转录聚合酶链反应(RT-PCR)评估病毒复制情况,同时在显微镜下观察细胞病变效应。使用下一代测序进行基因组测序,并评估瑞德西韦、莫努匹拉韦和奈玛特韦的抗病毒活性。
从一份儿童鼻拭子中成功分离出一株名为N15的SARS-CoV-2毒株。该分离株在Vero细胞中高效复制,其动力学与B.1和B.1.1.529谱系相当。感染后48小时内出现细胞病变效应,表现为死亡细胞聚集。基因组分析将N15归类为19B谱系,与武汉-1毒株的相似度为99.9%。鉴定出罕见突变:刺突蛋白中的N709S和包膜蛋白中的T11M。抗病毒测试显示瑞德西韦、莫努匹拉韦和奈玛特韦具有有效抑制作用,不同细胞类型的半数抑制浓度(IC50)值有所不同。
儿童N15分离株代表一种独特的19B谱系病毒,保留了祖先基因组特征,同时存在罕见突变。其高效复制和药物敏感性突出了其作为参考毒株的价值,可用于与流行变体进行比较研究以及评估抗病毒疗效。