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Here three complementary serological methods are described with different strengths to aid the development and evaluation of in\u2010house assays: An immunofluorescence assay (IFA) for specific detection of IgG and IgM, an enzyme\u2010linked immunosorbent assay for higher sample throughput to aid epidemiological studies and a neutralization test to detect virus neutralizing antibodies. As implementation of MPXV\u2010specific diagnostics is often hampered by the requirement for a dedicated biosafety level 3 laboratory (BSL\u20103), the focus of this study is on biosafety aspects to facilitate safe testing also under BSL\u20102 conditions. To this aim, it was analyzed whether OPXV, which can be handled under BSL\u20102 conditions, could be used as less virulent alternatives to MPXV. Furthermore, an inactivation method was established to remove up to five log\u2010steps of infectious virus particles from viraemic sera without compromising antibody detection. The results show that immunological cross\u2010reactivity between OPXV provides an opportunity for the interchangeable usage of different OPXV species in serological assays, enabling MPXV serology outside of BSL\u20103 facilities.<\/jats:p>","DOI":"10.1002\/jmv.29261","type":"journal-article","created":{"date-parts":[[2023,12,6]],"date-time":"2023-12-06T13:36:29Z","timestamp":1701869789000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":26,"title":["Serological methods for the detection of antibodies against monkeypox virus applicable for laboratories with different biosafety levels"],"prefix":"10.1002","volume":"95","author":[{"given":"Marica","family":"Grossegesse","sequence":"first","affiliation":[{"name":"Robert Koch Institute Centre for Biological Threats and Special Pathogens: Highly Pathogenic Viruses (ZBS 1)  Berlin Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniel","family":"Stern","sequence":"additional","affiliation":[{"name":"Robert Koch Institute Centre for Biological Threats and Special Pathogens: Biological Toxins (ZBS 3) Berlin Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Natalie","family":"Hofmann","sequence":"additional","affiliation":[{"name":"Robert Koch Institute Centre for Biological Threats and Special Pathogens: Highly Pathogenic Viruses (ZBS 1)  Berlin Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rebecca","family":"Surtees","sequence":"additional","affiliation":[{"name":"Robert Koch Institute Centre for Biological Threats and Special Pathogens: Highly Pathogenic Viruses (ZBS 1)  Berlin Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Claudia","family":"Kohl","sequence":"additional","affiliation":[{"name":"Robert Koch Institute Centre for Biological Threats and Special Pathogens: Highly Pathogenic Viruses (ZBS 1)  Berlin Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Janine","family":"Michel","sequence":"additional","affiliation":[{"name":"Robert Koch Institute Centre for Biological Threats and Special Pathogens: Highly Pathogenic Viruses (ZBS 1)  Berlin Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andreas","family":"Nitsche","sequence":"additional","affiliation":[{"name":"Robert Koch Institute Centre for Biological Threats and Special Pathogens: Highly Pathogenic Viruses (ZBS 1)  Berlin Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2023,12,6]]},"reference":[{"key":"e_1_2_9_2_1","first-page":"2333","volume-title":"Poxviridae: The viruses and their replication. 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