Luo Meihui, Shan Yuanchun, Zhang Xin, Ling Hua, Zhao Li, Huang Baoying, Wu Changcheng, Ruhan A, Deng Yao, Zhao Hua, Wang Wen, Ren Jiao, Ye Fei, Li Baisheng, Yang Xianda, Wang Huijuan, Huo Weibang, Zhai Yuqian, Han Yize, Tian Houwen, Lu Roujian, Wang Wenling, Tan Wenjie
National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, China.
Beijing Daxing District Center for Disease Prevention and Control, Beijing 102600, China.
Biosaf Health. 2025 Mar 22;7(2):103-109. doi: 10.1016/j.bsheal.2025.03.001. eCollection 2025 Apr.
In 2022, a global outbreak of mpox was anticipated, with several cases reported in non-endemic countries in early May. Given the challenge of distinguishing the mpox virus (MPXV) from other pathogens based solely on symptoms, there is an urgent need for prompt and reliable MPXV detection methods. In this study, we developed assays using recombinase-aided amplification (RAA) to identify MPXV and evaluated their applicability with clinical samples. The assays were designed to detect the gene of MPXV. All assays demonstrated detection limits of 1 copy/µL within the reaction system and exhibited no cross-reactivity with ectromelia or the TianTan strain of vaccinia virus, confirming their high specificity. Our established assay provides results in less than 50 min. Furthermore, we evaluated our assay using clinical samples from laboratory-confirmed mpox patients and demonstrated that the RAA-based assay is valuable for diagnosing MPXV infections in field and clinic settings, especially in areas with limited laboratory resources. Overall, three RAA-based nucleic acid assays for MPXV were established, providing a powerful tool for efficient, rapid, and specific detection of MPXV infection.
2022年，预计会出现猴痘全球大流行，5月初在非流行国家报告了多起病例。鉴于仅根据症状将猴痘病毒（MPXV）与其他病原体区分开来具有挑战性，因此迫切需要快速可靠的MPXV检测方法。在本研究中，我们开发了使用重组酶辅助扩增（RAA）的检测方法来鉴定MPXV，并评估了它们对临床样本的适用性。这些检测方法旨在检测MPXV的 基因。所有检测方法在反应体系中的检测限均为1拷贝/µL，并且与痘苗病毒的兔痘或天坛株无交叉反应，证实了它们的高特异性。我们建立的检测方法在不到50分钟内即可得出结果。此外，我们使用实验室确诊的猴痘患者的临床样本对检测方法进行了评估，结果表明基于RAA的检测方法对于在现场和临床环境中诊断MPXV感染非常有价值，尤其是在实验室资源有限的地区。总体而言，建立了三种基于RAA的MPXV核酸检测方法，为高效、快速和特异性检测MPXV感染提供了有力工具。
