{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,16]],"date-time":"2025-10-16T12:41:47Z","timestamp":1760618507203,"version":"build-2065373602"},"reference-count":11,"publisher":"Central Research Institute for Epidemiology","issue":"4","license":[{"start":{"date-parts":[[2025,9,16]],"date-time":"2025-09-16T00:00:00Z","timestamp":1757980800000},"content-version":"unspecified","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"content-domain":{"domain":["virusjour.crie.ru"],"crossmark-restriction":false},"short-container-title":["Problems of Virology"],"abstract":"<jats:p>Introduction. Highly pathogenic for humans orthopoxviruses (OPV) \u2013 Variola virus (VARV) and Monkeypox virus (MPXV) can cause systemic diseases characterized by high contagiousness and often leading to death. In previous publications (https:\/\/doi.org\/10.3390\/v14112580), we reported on the development of a sensitive, rapid and easy-to-use immunochemical test potentially suitable for use in the infection foci and in laboratory conditions with a high level of biosecurity. The prototype of the diagnostic kit was tested on non-pathogenic and low-pathogenic for humans OPV and specifically detected them with a sensitivity within the range of 103 to 104 PFU\/mL.\r\nThe aim of this work was to evaluate the sensitivity of this assay in detecting highly pathogenic for humans MPXV and VARV, as well as the applicability of the assay in a laboratory with a high level of biosafety (BSL-4).\r\nMaterials and methods. The efficiency of virus detection in cryolysates of CV-1 cell culture samples infected with VARV and MPXV was assessed using a one-step dot immunoassay method in a laboratory with biosafety level BSL-4.\r\nResults. It was shown that the one-step dot assay allows detection of MPXV at a concentration of 2.5 \u00d7 103 PFU\/mL, and VARV at a concentration of 1.0 \u00d7 104 PFU\/mL. It was noted that vapors from disinfectants (hydrogen peroxide\/formaldehyde) applied in biosafety cabinet decontamination may interfere with assay performance and affect result interpretation.\r\nConclusion. The one-step dot immunoassay can be performed in a BSL-4 laboratory. When limiting the contact of the detecting system with formaldehyde vapors, the sensitivity of the test for detection of VARV and MPXV falls within the previously declared range of 103\u2013104 PFU\/mL.<\/jats:p>","DOI":"10.36233\/0507-4088-332","type":"journal-article","created":{"date-parts":[[2025,9,16]],"date-time":"2025-09-16T18:36:20Z","timestamp":1758047780000},"page":"388-392","update-policy":"https:\/\/doi.org\/10.36233\/0372-9311-crossmark","source":"Crossref","is-referenced-by-count":0,"title":["Detection of highly pathogenic orthopoxviruses (&lt;i&gt;Poxviridae&lt;\/i&gt;: &lt;i&gt;Chordopoxvirinae&lt;\/i&gt;: &lt;i&gt;Orthopoxvirus&lt;\/i&gt;) by dot immunoassay in a high-containment biological safety laboratory"],"prefix":"10.36233","volume":"70","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9220-1250","authenticated-orcid":false,"given":"Anna V.","family":"Ersh","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2408-5611","authenticated-orcid":false,"given":"Alexander G.","family":"Poltavchenko","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2171-7444","authenticated-orcid":false,"given":"Nikita D.","family":"Ushkalenko","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7763-3808","authenticated-orcid":false,"given":"Pavel V.","family":"Filatov","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8764-9408","authenticated-orcid":false,"given":"Ksenia A.","family":"Titova","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0616-7488","authenticated-orcid":false,"given":"Dmitrii A.","family":"Odnoshevsky","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8355-5551","authenticated-orcid":false,"given":"Alexander A.","family":"Sergeev","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3591-1571","authenticated-orcid":false,"given":"Artemiy A.","family":"Sergeev","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"22004","published-online":{"date-parts":[[2025,9,16]]},"reference":[{"key":"ref1","doi-asserted-by":"crossref","unstructured":"Buller R.M.L. Poxviruses. In: Cohen J., Powderly W.G., Opal S.M., eds. Infectious Diseases. Volume 2. Elsevier; 2017: 1452\u20137.e1.","DOI":"10.1016\/B978-0-7020-6285-8.00170-2"},{"key":"ref2","doi-asserted-by":"crossref","unstructured":"Shchelkunova G.A., Shchelkunov S.N. Smallpox, monkeypox and other human orthopoxvirus infections. Viruses. 2022; 15(1): 103. https:\/\/doi.org\/10.3390\/v15010103","DOI":"10.3390\/v15010103"},{"key":"ref3","doi-asserted-by":"crossref","unstructured":"Sklenovsk\u00e1 N. Monkeypox virus. In: Malik Y.S., Singh R.K., Dhama K., eds. Livestock Diseases and Management. Springer; 2020: 39\u201368.","DOI":"10.1007\/978-981-15-2651-0_2"},{"key":"ref4","doi-asserted-by":"crossref","unstructured":"Whitley R.J. Smallpox: a potential agent of bioterrorism. Antiviral. Res. 2003; 57(1-2): 7\u201312. https:\/\/doi.org\/10.1016\/s0166-3542(02)00195-x","DOI":"10.1016\/S0166-3542(02)00195-X"},{"key":"ref5","doi-asserted-by":"crossref","unstructured":"Wallin A., Luksiene Z., Zagminas K., Surkiene G. Public health and bioterrorism: renewed threat of anthrax and smallpox. Medicina (Kaunas). 2007; 43(4): 278\u201384.","DOI":"10.3390\/medicina43040034"},{"key":"ref6","doi-asserted-by":"crossref","unstructured":"Harapan H., Ophinni Y., Megawati D., Frediansyah A., Mamada S.S., Salampe M., et al. Monkeypox: a comprehensive review. Viruses. 2022; 14(10): 2155. https:\/\/doi.org\/10.3390\/v14102155","DOI":"10.3390\/v14102155"},{"key":"ref7","doi-asserted-by":"crossref","unstructured":"Shurtleff A.C., Garza N., Lackemeyer M., Carrion R. Jr., Griffiths A., Patterson J., et al. The impact of regulations, safety considerations and physical limitations on research progress at maximum biocontainment. Viruses. 2012; 4(12): 3932\u201351. https:\/\/doi.org\/10.3390\/v4123932","DOI":"10.3390\/v4123932"},{"key":"ref8","doi-asserted-by":"crossref","unstructured":"Bohannon J.K., Janosko K., Holbrook M.R., Barr J., Pusl D., Bollinger L., et al. Safety precautions and operating procedures in an (A)BSL-4 laboratory: 3. Aerobiology. J. Vis. Exp. 2016; (116): 53602. https:\/\/doi.org\/10.3791\/53602","DOI":"10.3791\/53602-v"},{"key":"ref9","doi-asserted-by":"crossref","unstructured":"Townsend M.B., MacNeil A., Reynolds M.G., Hughes C.M., Olson V.A., Damon I.K., et al. Evaluation of the Tetracore Orthopox BioThreat\u00ae antigen detection assay using laboratory grown orthopoxviruses and rash illness clinical specimens. J. Virol. Methods. 2013; 187(1): 37\u201342. https:\/\/doi.org\/10.1016\/j.jviromet.2012.08.023","DOI":"10.1016\/j.jviromet.2012.08.023"},{"key":"ref10","doi-asserted-by":"crossref","unstructured":"Stern D., Olson V.A., Smith S.K., Pietraszczyk M., Miller L., Miethe P., et al. Rapid and sensitive point-of-care detection of Orthopoxviruses by ABICAP immunofiltration. Virol. J. 2016; 13(1): 207. https:\/\/doi.org\/10.1186\/s12985-016-0665-5","DOI":"10.1186\/s12985-016-0665-5"},{"key":"ref11","doi-asserted-by":"crossref","unstructured":"Ushkalenko N., Ersh A., Sergeev A., Filatov P., Poltavchenko A. Evaluation of rapid dot-immunoassay for detection orthopoxviruses using laboratory-grown viruses and animal\u2019s clinical specimens. Viruses. 2022; 14(11): 2580. https:\/\/doi.org\/10.3390\/v14112580","DOI":"10.3390\/v14112580"}],"container-title":["Problems of Virology"],"original-title":[],"link":[{"URL":"https:\/\/virusjour.crie.ru\/jour\/article\/viewFile\/16768\/1008","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/virusjour.crie.ru\/jour\/article\/viewFile\/16768\/1008","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,16]],"date-time":"2025-10-16T12:21:50Z","timestamp":1760617310000},"score":1,"resource":{"primary":{"URL":"https:\/\/virusjour.crie.ru\/jour\/article\/view\/16768"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,9,16]]},"references-count":11,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2025,9,16]]}},"URL":"https:\/\/doi.org\/10.36233\/0507-4088-332","relation":{},"ISSN":["2411-2097","0507-4088"],"issn-type":[{"type":"electronic","value":"2411-2097"},{"type":"print","value":"0507-4088"}],"subject":[],"published":{"date-parts":[[2025,9,16]]}}}