Extraction-free LAMP assays for generic detection of Old World Orthopoxviruses and specific detection of Mpox virus
Abstract Mpox is a neglected zoonotic disease endemic in West and Central Africa. The Mpox outbreak with more than 90,000 cases worldwide since 2022 generated great concern about future outbreaks and highlighted the need for a simple and rapid diagnostic test. The Mpox virus, MPV, is a member of the...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2023-11-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-023-48391-z |
| _version_ | 1797415169019609088 |
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| author | Zhiru Li Amit Sinha Yinhua Zhang Nathan Tanner Hui-Ting Cheng Prem Premsrirut Clotilde K. S. Carlow |
| author_facet | Zhiru Li Amit Sinha Yinhua Zhang Nathan Tanner Hui-Ting Cheng Prem Premsrirut Clotilde K. S. Carlow |
| author_sort | Zhiru Li |
| collection | DOAJ |
| description | Abstract Mpox is a neglected zoonotic disease endemic in West and Central Africa. The Mpox outbreak with more than 90,000 cases worldwide since 2022 generated great concern about future outbreaks and highlighted the need for a simple and rapid diagnostic test. The Mpox virus, MPV, is a member of the Orthopoxvirus (OPV) genus that also contains other pathogenic viruses including variola virus, vaccinia virus, camelpox virus, and cowpox virus. Phylogenomic analysis of 200 OPV genomes identified 10 distinct phylogroups with the New World OPVs placed on a very long branch distant from the Old World OPVs. Isolates derived from infected humans were found to be distributed across multiple phylogroups interspersed with isolates from animal sources, indicating the zoonotic potential of these viruses. In this study, we developed a simple and sensitive colorimetric LAMP assay for generic detection of Old World OPVs. We also developed an MPV-specific probe that differentiates MPV from other OPVs in the N1R LAMP assay. In addition, we described an extraction-free protocol for use directly with swab eluates in LAMP assays, thereby eliminating the time and resources needed to extract DNA from the sample. Our direct LAMP assays are well-suited for low-resource settings and provide a valuable tool for rapid and scalable diagnosis and surveillance of OPVs and MPV. |
| first_indexed | 2024-03-09T05:43:58Z |
| format | Article |
| id | doaj.art-433140d545d2457c9b9285c1fa08806f |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-03-09T05:43:58Z |
| publishDate | 2023-11-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-433140d545d2457c9b9285c1fa08806f2023-12-03T12:22:38ZengNature PortfolioScientific Reports2045-23222023-11-0113111210.1038/s41598-023-48391-zExtraction-free LAMP assays for generic detection of Old World Orthopoxviruses and specific detection of Mpox virusZhiru Li0Amit Sinha1Yinhua Zhang2Nathan Tanner3Hui-Ting Cheng4Prem Premsrirut5Clotilde K. S. Carlow6Molecular Genetics and Genomics Division, New England BiolabsMolecular Genetics and Genomics Division, New England BiolabsMolecular Genetics and Genomics Division, New England BiolabsMolecular Genetics and Genomics Division, New England BiolabsMirimus Inc.Mirimus Inc.Molecular Genetics and Genomics Division, New England BiolabsAbstract Mpox is a neglected zoonotic disease endemic in West and Central Africa. The Mpox outbreak with more than 90,000 cases worldwide since 2022 generated great concern about future outbreaks and highlighted the need for a simple and rapid diagnostic test. The Mpox virus, MPV, is a member of the Orthopoxvirus (OPV) genus that also contains other pathogenic viruses including variola virus, vaccinia virus, camelpox virus, and cowpox virus. Phylogenomic analysis of 200 OPV genomes identified 10 distinct phylogroups with the New World OPVs placed on a very long branch distant from the Old World OPVs. Isolates derived from infected humans were found to be distributed across multiple phylogroups interspersed with isolates from animal sources, indicating the zoonotic potential of these viruses. In this study, we developed a simple and sensitive colorimetric LAMP assay for generic detection of Old World OPVs. We also developed an MPV-specific probe that differentiates MPV from other OPVs in the N1R LAMP assay. In addition, we described an extraction-free protocol for use directly with swab eluates in LAMP assays, thereby eliminating the time and resources needed to extract DNA from the sample. Our direct LAMP assays are well-suited for low-resource settings and provide a valuable tool for rapid and scalable diagnosis and surveillance of OPVs and MPV.https://doi.org/10.1038/s41598-023-48391-z |
| spellingShingle | Zhiru Li Amit Sinha Yinhua Zhang Nathan Tanner Hui-Ting Cheng Prem Premsrirut Clotilde K. S. Carlow Extraction-free LAMP assays for generic detection of Old World Orthopoxviruses and specific detection of Mpox virus Scientific Reports |
| title | Extraction-free LAMP assays for generic detection of Old World Orthopoxviruses and specific detection of Mpox virus |
| title_full | Extraction-free LAMP assays for generic detection of Old World Orthopoxviruses and specific detection of Mpox virus |
| title_fullStr | Extraction-free LAMP assays for generic detection of Old World Orthopoxviruses and specific detection of Mpox virus |
| title_full_unstemmed | Extraction-free LAMP assays for generic detection of Old World Orthopoxviruses and specific detection of Mpox virus |
| title_short | Extraction-free LAMP assays for generic detection of Old World Orthopoxviruses and specific detection of Mpox virus |
| title_sort | extraction free lamp assays for generic detection of old world orthopoxviruses and specific detection of mpox virus |
| url | https://doi.org/10.1038/s41598-023-48391-z |
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