由于缺乏有效的诊断工具，区分EBV阳性患者的良性和恶性疾病是一个重大挑战。元基因组的新一代测序（mNGS）通常用于鉴定原因不明热患者的病原体。最近的研究已经扩展了新一代测序（NGS）在体液和脑脊液中鉴定肿瘤的应用。基于这些研究，我们进行了这个研究，旨在开发和应用元基因组学方法来验证其在识别EBV相关恶性疾病中的作用。 我们选择2018年至2019年期间在复旦大学附属华山医院感染科发热原因不明患者中被检测出EBV阳性的29例患者作为研究对象。在入组时，根据最终诊断将这些患者分为良性疾病、慢性活动性EBV感染和恶性疾病组，并于2022年对2018年至2019年的样本进行回顾性CNV分析。 在29名患者中，16名被诊断为良性疾病，3名被诊断为慢性活动性EBV感染，10名被诊断为恶性疾病。我们对这29名患者的血样进行了mNGS测试。在所有被诊断为恶性疾病的10名患者中，CNV分析提示有9名患者存在肿瘤。在19名被诊断为良性或慢性活动性EBV感染的患者中，有2名患者的CNV结果异常。CNV分析在肿瘤识别中的敏感性和特异性分别为90％和89.5％。 mNGS的应用可以帮助鉴定EBV相关疾病中的微生物感染和恶性肿瘤。我们的研究结果表明，通过mNGS进行CNV检测比传统的肿瘤学检测方法更快。此外，外周血样本的便捷采集也是这种方法的优势所在。 版权所有 © 2023 Song, Zhu, Wang, Yang, Yu, Zhang, Fu, Wang, Zhao, Lin, Yuan, Guo, Shi, Liu, Ai, Zhang and Zhang.

Differentiation between benign and malignant diseases in EBV-positive patients poses a significant challenge due to the lack of efficient diagnostic tools. Metagenomic Next-Generation Sequencing (mNGS) is commonly used to identify pathogens of patients with fevers of unknown-origin (FUO). Recent studies have extended the application of Next-Generation Sequencing (NGS) in identifying tumors in body fluids and cerebrospinal fluids. In light of these, we conducted this study to develop and apply metagenomic methods to validate their role in identifying EBV-associated malignant disease.We enrolled 29 patients with positive EBV results in the cohort of FUO in the Department of Infectious Diseases of Huashan Hospital affiliated with Fudan University from 2018 to 2019. Upon enrollment, these patients were grouped for benign diseases, CAEBV, and malignant diseases according to their final diagnosis, and CNV analysis was retrospectively performed in 2022 using samples from 2018 to 2019.Among the 29 patients. 16 of them were diagnosed with benign diseases, 3 patients were diagnosed with CAEBV and 10 patients were with malignant diseases. 29 blood samples from 29 patients were tested for mNGS. Among all 10 patients with malignant diagnosis, CNV analysis suggested neoplasms in 9 patients. Of all 19 patients with benign or CAEBV diagnosis, 2 patients showed abnormal CNV results. The sensitivity and specificity of CNV analysis for the identification for tumors were 90% and 89.5%, separately.The application of mNGS could assist in the identification of microbial infection and malignancies in EBV-related diseases. Our results demonstrate that CNV detection through mNGS is faster compared to conventional oncology tests. Moreover, the convenient collection of peripheral blood samples adds to the advantages of this approach.Copyright © 2023 Song, Zhu, Wang, Yang, Yu, Zhang, Fu, Wang, Zhao, Lin, Yuan, Guo, Shi, Liu, Ai, Zhang and Zhang.