Comprehensive Assessment of Inactivation Methods for Madariaga Virus
The Eastern Equine Encephalitis Virus (EEEV) is an emerging public health threat, with the number of reported cases in the US increasing in recent years. EEEV is a BSL3 pathogen, and the North American strain is a US Federal Select Agent (SA). These restrictions make experiments with EEEV difficult...
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| Format: | Article |
| Language: | English |
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MDPI AG
2024-01-01
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| Series: | Viruses |
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| Online Access: | https://www.mdpi.com/1999-4915/16/2/206 |
| _version_ | 1797296809250390016 |
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| author | RuthMabel Boytz Kadiatou Keita Joanna B Pawlak Maudry Laurent-Rolle |
| author_facet | RuthMabel Boytz Kadiatou Keita Joanna B Pawlak Maudry Laurent-Rolle |
| author_sort | RuthMabel Boytz |
| collection | DOAJ |
| description | The Eastern Equine Encephalitis Virus (EEEV) is an emerging public health threat, with the number of reported cases in the US increasing in recent years. EEEV is a BSL3 pathogen, and the North American strain is a US Federal Select Agent (SA). These restrictions make experiments with EEEV difficult to perform, as high-tech equipment is often unavailable in BSL3 spaces and due to concerns about generating aerosols during manipulations. Therefore, a range of inactivation methods suitable for different downstream analysis methods are essential for advancing research on EEEV. We used heat, chemical, and ultraviolet (UV)-based methods for the inactivation of infected cells and supernatants infected with the non-select agent Madariaga virus (MADV). Although the MADV and EEEV strains are genetically distinct, differing by 8–11% at the amino acid level, they are expected to be similarly susceptible to various inactivation methods. We determined the following to be effective methods of inactivation: heat, TRIzol LS, 4% PFA, 10% formalin, and UV radiation for infected supernatants; TRIzol, 2.5% SDS with BME, 0.2% NP40, 4% PFA, and 10% formalin for infected cells. Our results have the potential to expand the types and complexity of experiments and analyses performed by EEEV researchers. |
| first_indexed | 2024-03-07T22:11:06Z |
| format | Article |
| id | doaj.art-737ac87e83a342e7804f29d206d1538f |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-07T22:11:06Z |
| publishDate | 2024-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj.art-737ac87e83a342e7804f29d206d1538f2024-02-23T15:37:29ZengMDPI AGViruses1999-49152024-01-0116220610.3390/v16020206Comprehensive Assessment of Inactivation Methods for Madariaga VirusRuthMabel Boytz0Kadiatou Keita1Joanna B Pawlak2Maudry Laurent-Rolle3Department of Microbial Pathogenesis, Yale University School of Medicine, New Haven, CT 06520, USASection of Infectious Diseases, Department of Internal Medicine, Yale University School of Medicine, New Haven, CT 06520, USASection of Infectious Diseases, Department of Internal Medicine, Yale University School of Medicine, New Haven, CT 06520, USADepartment of Microbial Pathogenesis, Yale University School of Medicine, New Haven, CT 06520, USAThe Eastern Equine Encephalitis Virus (EEEV) is an emerging public health threat, with the number of reported cases in the US increasing in recent years. EEEV is a BSL3 pathogen, and the North American strain is a US Federal Select Agent (SA). These restrictions make experiments with EEEV difficult to perform, as high-tech equipment is often unavailable in BSL3 spaces and due to concerns about generating aerosols during manipulations. Therefore, a range of inactivation methods suitable for different downstream analysis methods are essential for advancing research on EEEV. We used heat, chemical, and ultraviolet (UV)-based methods for the inactivation of infected cells and supernatants infected with the non-select agent Madariaga virus (MADV). Although the MADV and EEEV strains are genetically distinct, differing by 8–11% at the amino acid level, they are expected to be similarly susceptible to various inactivation methods. We determined the following to be effective methods of inactivation: heat, TRIzol LS, 4% PFA, 10% formalin, and UV radiation for infected supernatants; TRIzol, 2.5% SDS with BME, 0.2% NP40, 4% PFA, and 10% formalin for infected cells. Our results have the potential to expand the types and complexity of experiments and analyses performed by EEEV researchers.https://www.mdpi.com/1999-4915/16/2/206MadariagaEastern Equine Encephalitis VirusinactivationprotocolSelect Agent |
| spellingShingle | RuthMabel Boytz Kadiatou Keita Joanna B Pawlak Maudry Laurent-Rolle Comprehensive Assessment of Inactivation Methods for Madariaga Virus Viruses Madariaga Eastern Equine Encephalitis Virus inactivation protocol Select Agent |
| title | Comprehensive Assessment of Inactivation Methods for Madariaga Virus |
| title_full | Comprehensive Assessment of Inactivation Methods for Madariaga Virus |
| title_fullStr | Comprehensive Assessment of Inactivation Methods for Madariaga Virus |
| title_full_unstemmed | Comprehensive Assessment of Inactivation Methods for Madariaga Virus |
| title_short | Comprehensive Assessment of Inactivation Methods for Madariaga Virus |
| title_sort | comprehensive assessment of inactivation methods for madariaga virus |
| topic | Madariaga Eastern Equine Encephalitis Virus inactivation protocol Select Agent |
| url | https://www.mdpi.com/1999-4915/16/2/206 |
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