Detection of the mpox virus using a robust recombinase-aided amplification-based approach

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...

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Main Authors: Meihui Luo, Yuanchun Shan, Xin Zhang, Hua Ling, Li Zhao, Baoying Huang, Changcheng Wu, A Ruhan, Yao Deng, Hua Zhao, Wen Wang, Jiao Ren, Fei Ye, Baisheng Li, Xianda Yang, Huijuan Wang, Weibang Huo, Yuqian Zhai, Yize Han, Houwen Tian, Roujian Lu, Wenling Wang, Wenjie Tan
Format: Article
Language:English
Published: Elsevier 2025-04-01
Series:Biosafety and Health
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Online Access:http://www.sciencedirect.com/science/article/pii/S2590053625000370
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author Meihui Luo
Yuanchun Shan
Xin Zhang
Hua Ling
Li Zhao
Baoying Huang
Changcheng Wu
A Ruhan
Yao Deng
Hua Zhao
Wen Wang
Jiao Ren
Fei Ye
Baisheng Li
Xianda Yang
Huijuan Wang
Weibang Huo
Yuqian Zhai
Yize Han
Houwen Tian
Roujian Lu
Wenling Wang
Wenjie Tan
author_facet Meihui Luo
Yuanchun Shan
Xin Zhang
Hua Ling
Li Zhao
Baoying Huang
Changcheng Wu
A Ruhan
Yao Deng
Hua Zhao
Wen Wang
Jiao Ren
Fei Ye
Baisheng Li
Xianda Yang
Huijuan Wang
Weibang Huo
Yuqian Zhai
Yize Han
Houwen Tian
Roujian Lu
Wenling Wang
Wenjie Tan
author_sort Meihui Luo
collection DOAJ
description 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 N4R 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.
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spelling doaj-art-1587f84f3f594eebbb8bc5e72bbfbfba2025-08-20T03:13:53ZengElsevierBiosafety and Health2590-05362025-04-017210310910.1016/j.bsheal.2025.03.001Detection of the mpox virus using a robust recombinase-aided amplification-based approachMeihui Luo0Yuanchun Shan1Xin Zhang2Hua Ling3Li Zhao4Baoying Huang5Changcheng Wu6A Ruhan7Yao Deng8Hua Zhao9Wen Wang10Jiao Ren11Fei Ye12Baisheng Li13Xianda Yang14Huijuan Wang15Weibang Huo16Yuqian Zhai17Yize Han18Houwen Tian19Roujian Lu20Wenling Wang21Wenjie Tan22National 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, ChinaJilin Center for Disease Control and Prevention, Jilin 130062,ChinaGuangdong Provincial Center for Disease Control and Prevention, Guangzhou 511430, ChinaChongqing Center for Disease Control and Prevention, Chongqing 400042, ChinaNational Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, ChinaNational Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, ChinaNational Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, ChinaNational Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, ChinaNational Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, ChinaJilin Center for Disease Control and Prevention, Jilin 130062,ChinaNational Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, ChinaNational Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, ChinaNational Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, ChinaGuangdong Provincial Center for Disease Control and Prevention, Guangzhou 511430, ChinaJilin Center for Disease Control and Prevention, Jilin 130062,ChinaNational Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, ChinaNational Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, ChinaNational Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, ChinaNational Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, ChinaNational Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, ChinaNational Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, ChinaNational Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, China; Corresponding author: NHC Key Laboratory of Biosafety, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, 155 Changbai Road, Changping District, Beijing 102206, China.National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, ChinaIn 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 N4R 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.http://www.sciencedirect.com/science/article/pii/S2590053625000370Mpox virusNucleic acid detectionRecombinase-aided amplification (RAA)Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated 12a (Cas12a)
spellingShingle Meihui Luo
Yuanchun Shan
Xin Zhang
Hua Ling
Li Zhao
Baoying Huang
Changcheng Wu
A Ruhan
Yao Deng
Hua Zhao
Wen Wang
Jiao Ren
Fei Ye
Baisheng Li
Xianda Yang
Huijuan Wang
Weibang Huo
Yuqian Zhai
Yize Han
Houwen Tian
Roujian Lu
Wenling Wang
Wenjie Tan
Detection of the mpox virus using a robust recombinase-aided amplification-based approach
Biosafety and Health
Mpox virus
Nucleic acid detection
Recombinase-aided amplification (RAA)
Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated 12a (Cas12a)
title Detection of the mpox virus using a robust recombinase-aided amplification-based approach
title_full Detection of the mpox virus using a robust recombinase-aided amplification-based approach
title_fullStr Detection of the mpox virus using a robust recombinase-aided amplification-based approach
title_full_unstemmed Detection of the mpox virus using a robust recombinase-aided amplification-based approach
title_short Detection of the mpox virus using a robust recombinase-aided amplification-based approach
title_sort detection of the mpox virus using a robust recombinase aided amplification based approach
topic Mpox virus
Nucleic acid detection
Recombinase-aided amplification (RAA)
Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated 12a (Cas12a)
url http://www.sciencedirect.com/science/article/pii/S2590053625000370
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