Rapid detection of mpox virus using recombinase aided amplification assay
A recent, unprecedented outbreak of human mpox virus infection has led to cases in non-African nations, and the number of confirmed or suspected cases outside of Africa has exceeded 1,000 within 5 weeks. Mpox may pose a double threat to public health in the context of the ongoing COVID-19 pandemic....
| Main Authors: | , , , , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2023-02-01
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| Series: | Frontiers in Cellular and Infection Microbiology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fcimb.2023.1008783/full |
| _version_ | 1797896546466922496 |
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| author | Xiaohu Cui Bing Du Bing Du Junxia Feng Yanling Feng Jinghua Cui Chao Yan Hanqing Zhao Lin Gan Zheng Fan Tongtong Fu Ziying Xu Rui Zhang Shuheng Du Yao Zhou Ziyan Tian Qun Zhang Hanyu Fu Guanhua Xue Jing Yuan |
| author_facet | Xiaohu Cui Bing Du Bing Du Junxia Feng Yanling Feng Jinghua Cui Chao Yan Hanqing Zhao Lin Gan Zheng Fan Tongtong Fu Ziying Xu Rui Zhang Shuheng Du Yao Zhou Ziyan Tian Qun Zhang Hanyu Fu Guanhua Xue Jing Yuan |
| author_sort | Xiaohu Cui |
| collection | DOAJ |
| description | A recent, unprecedented outbreak of human mpox virus infection has led to cases in non-African nations, and the number of confirmed or suspected cases outside of Africa has exceeded 1,000 within 5 weeks. Mpox may pose a double threat to public health in the context of the ongoing COVID-19 pandemic. It is difficult to distinguish mpox virus infection from other diseases in the early stages, and patients are contagious from the onset of nonspecific symptoms; therefore, it is crucial to develop rapid and specific diagnostic methods. The diagnosis of mpox relies on real-time polymerase chain reaction, a time-consuming method that requires a highly sophisticated thermal cycler, which makes it unsuitable for widespread use in underdeveloped areas, where the outbreak is still severe. In this study, we developed a recombinase-aided amplification (RAA) assay that can detect mpox virus within 5–10 minutes. The conserved regions of the A27L gene and F3L gene were selected as targets, as they amplify well from different mpox virus clades with no cross-reaction from other pathogens. The sensitivity of this RAA assay is 10 copies/reaction for the A27L gene and 102 copies/reaction for the F3L gene. When applied to simulated clinical samples, both targets showed 100% specificity, and the detection limits were consistent with the sensitivity results. Moreover, through clinical blinded sample detection, RAA exhibits the same detection power as RT-PCR. In summary, the RAA mpox assay described here exhibits rapid detection, high sensitivity and specificity, and low operational difficulty, making it suitable for mpox virus detection in less developed countries and regions. |
| first_indexed | 2024-04-10T07:44:34Z |
| format | Article |
| id | doaj.art-8095699a08e8476aa2dac016484bdeb5 |
| institution | Directory Open Access Journal |
| issn | 2235-2988 |
| language | English |
| last_indexed | 2024-04-10T07:44:34Z |
| publishDate | 2023-02-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cellular and Infection Microbiology |
| spelling | doaj.art-8095699a08e8476aa2dac016484bdeb52023-02-23T11:04:45ZengFrontiers Media S.A.Frontiers in Cellular and Infection Microbiology2235-29882023-02-011310.3389/fcimb.2023.10087831008783Rapid detection of mpox virus using recombinase aided amplification assayXiaohu Cui0Bing Du1Bing Du2Junxia Feng3Yanling Feng4Jinghua Cui5Chao Yan6Hanqing Zhao7Lin Gan8Zheng Fan9Tongtong Fu10Ziying Xu11Rui Zhang12Shuheng Du13Yao Zhou14Ziyan Tian15Qun Zhang16Hanyu Fu17Guanhua Xue18Jing Yuan19Department of Bacteriology, Capital Institute of Pediatrics, Beijing, ChinaDepartment of Bacteriology, Capital Institute of Pediatrics, Beijing, ChinaSchool of Biological Sciences, The University of Edinburgh, Edinburgh, United KingdomDepartment of Bacteriology, Capital Institute of Pediatrics, Beijing, ChinaDepartment of Bacteriology, Capital Institute of Pediatrics, Beijing, ChinaDepartment of Bacteriology, Capital Institute of Pediatrics, Beijing, ChinaDepartment of Bacteriology, Capital Institute of Pediatrics, Beijing, ChinaDepartment of Bacteriology, Capital Institute of Pediatrics, Beijing, ChinaDepartment of Bacteriology, Capital Institute of Pediatrics, Beijing, ChinaDepartment of Bacteriology, Capital Institute of Pediatrics, Beijing, ChinaDepartment of Bacteriology, Capital Institute of Pediatrics, Beijing, ChinaDepartment of Bacteriology, Capital Institute of Pediatrics, Beijing, ChinaDepartment of Bacteriology, Capital Institute of Pediatrics, Beijing, ChinaDepartment of Bacteriology, Capital Institute of Pediatrics, Beijing, ChinaDepartment of Bacteriology, Capital Institute of Pediatrics, Beijing, ChinaDepartment of Bacteriology, Capital Institute of Pediatrics, Beijing, ChinaDepartment of Bacteriology, Capital Institute of Pediatrics, Beijing, ChinaDepartment of Bacteriology, Capital Institute of Pediatrics, Beijing, ChinaDepartment of Bacteriology, Capital Institute of Pediatrics, Beijing, ChinaDepartment of Bacteriology, Capital Institute of Pediatrics, Beijing, ChinaA recent, unprecedented outbreak of human mpox virus infection has led to cases in non-African nations, and the number of confirmed or suspected cases outside of Africa has exceeded 1,000 within 5 weeks. Mpox may pose a double threat to public health in the context of the ongoing COVID-19 pandemic. It is difficult to distinguish mpox virus infection from other diseases in the early stages, and patients are contagious from the onset of nonspecific symptoms; therefore, it is crucial to develop rapid and specific diagnostic methods. The diagnosis of mpox relies on real-time polymerase chain reaction, a time-consuming method that requires a highly sophisticated thermal cycler, which makes it unsuitable for widespread use in underdeveloped areas, where the outbreak is still severe. In this study, we developed a recombinase-aided amplification (RAA) assay that can detect mpox virus within 5–10 minutes. The conserved regions of the A27L gene and F3L gene were selected as targets, as they amplify well from different mpox virus clades with no cross-reaction from other pathogens. The sensitivity of this RAA assay is 10 copies/reaction for the A27L gene and 102 copies/reaction for the F3L gene. When applied to simulated clinical samples, both targets showed 100% specificity, and the detection limits were consistent with the sensitivity results. Moreover, through clinical blinded sample detection, RAA exhibits the same detection power as RT-PCR. In summary, the RAA mpox assay described here exhibits rapid detection, high sensitivity and specificity, and low operational difficulty, making it suitable for mpox virus detection in less developed countries and regions.https://www.frontiersin.org/articles/10.3389/fcimb.2023.1008783/fullMpox virusmolecular diagnostic methodrecombinase-aided amplificationRAA assayrapid detection |
| spellingShingle | Xiaohu Cui Bing Du Bing Du Junxia Feng Yanling Feng Jinghua Cui Chao Yan Hanqing Zhao Lin Gan Zheng Fan Tongtong Fu Ziying Xu Rui Zhang Shuheng Du Yao Zhou Ziyan Tian Qun Zhang Hanyu Fu Guanhua Xue Jing Yuan Rapid detection of mpox virus using recombinase aided amplification assay Frontiers in Cellular and Infection Microbiology Mpox virus molecular diagnostic method recombinase-aided amplification RAA assay rapid detection |
| title | Rapid detection of mpox virus using recombinase aided amplification assay |
| title_full | Rapid detection of mpox virus using recombinase aided amplification assay |
| title_fullStr | Rapid detection of mpox virus using recombinase aided amplification assay |
| title_full_unstemmed | Rapid detection of mpox virus using recombinase aided amplification assay |
| title_short | Rapid detection of mpox virus using recombinase aided amplification assay |
| title_sort | rapid detection of mpox virus using recombinase aided amplification assay |
| topic | Mpox virus molecular diagnostic method recombinase-aided amplification RAA assay rapid detection |
| url | https://www.frontiersin.org/articles/10.3389/fcimb.2023.1008783/full |
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