{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,23]],"date-time":"2026-09-23T13:00:35Z","timestamp":1790168435780,"version":"4.1.0"},"reference-count":34,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2022,12,21]],"date-time":"2022-12-21T00:00:00Z","timestamp":1671580800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2022,12,21]],"date-time":"2022-12-21T00:00:00Z","timestamp":1671580800000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/100007130","name":"University of Texas System","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100007130","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Journal of Medical Virology"],"published-print":{"date-parts":[[2023,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Rapid and accurate diagnosis of infections is fundamental to containment of disease. Several monkeypox virus (MPV) real\u2010time diagnostic assays have been recommended by the CDC; however, the specificity of the primers and probes in these assays for the ongoing MPV outbreak has not been investigated. We analyzed the primer and probe sequences present in the CDC recommended MPV\u00a0generic real\u2010time PCR assay by aligning those sequences against 1730 MPV complete genomes reported in 2022 worldwide. Sequence mismatches were found in 99.08% and 97.46% of genomes for the MPV generic forward and reverse primers, respectively. Mismatch\u2010corrected primers were synthetized and compared to the generic assay for MPV detection. Results showed that the two primer\u2010template mismatches resulted in a ~11\u2010fold underestimation of initial template DNA in the reaction and 4\u2010fold increase in the 95% LOD. We further evaluated the specificity of seven other real\u2010time PCR assays used for MPV and orthopoxvirus (OPV) detection and identified two assays with the highest matching score (&gt;99.6%) to the global MPV genome database in 2022. Genetic variations in the primer\u2010probe regions across MPV genomes could indicate the temporal and spatial emergence pattern of monkeypox disease. Our results show that the current MPV real\u2010time generic assay may not be optimal to accurately detect MPV, and the mismatch\u2010corrected assay with full complementarity between primers and current MPV genomes could provide a more sensitive and accurate detection of MPV.<\/jats:p>","DOI":"10.1002\/jmv.28395","type":"journal-article","created":{"date-parts":[[2022,12,12]],"date-time":"2022-12-12T04:24:03Z","timestamp":1670819043000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":26,"title":["Wide mismatches in the sequences of primers and probes for monkeypox virus diagnostic assays"],"prefix":"10.1002","volume":"95","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2820-3550","authenticated-orcid":false,"given":"Fuqing","family":"Wu","sequence":"first","affiliation":[{"name":"School of Public Health, The University of Texas Health Science Center at Houston Houston Texas USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jeremiah","family":"Oghuan","sequence":"additional","affiliation":[{"name":"School of Public Health, The University of Texas Health Science Center at Houston Houston Texas USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anna","family":"Gitter","sequence":"additional","affiliation":[{"name":"School of Public Health, The University of Texas Health Science Center at Houston Houston Texas USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kristina D.","family":"Mena","sequence":"additional","affiliation":[{"name":"School of Public Health, The University of Texas Health Science Center at Houston Houston Texas USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eric L.","family":"Brown","sequence":"additional","affiliation":[{"name":"School of Public Health, The University of Texas Health Science Center at Houston Houston Texas USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2022,12,21]]},"reference":[{"key":"e_1_2_9_2_1","volume-title":"Monkeypox Outbreak Global Map","author":"CDC.\u00a0","year":"2022"},{"key":"e_1_2_9_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.idc.2019.03.001"},{"key":"e_1_2_9_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/S1473-3099(21)00170-5"},{"issue":"2","key":"e_1_2_9_5_1","first-page":"165","article-title":"Human monkeypox, 1970\u201079","volume":"58","author":"Breman JG","year":"1980","journal-title":"Bull World Health Organ"},{"key":"e_1_2_9_6_1","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa2207323"},{"key":"e_1_2_9_7_1","doi-asserted-by":"publisher","DOI":"10.1093\/ofid\/ofac310"},{"key":"e_1_2_9_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jaut.2022.102855"},{"key":"e_1_2_9_9_1","doi-asserted-by":"publisher","DOI":"10.2807\/1560-7917.ES.2022.27.22.2200422"},{"key":"e_1_2_9_10_1","volume-title":"Test\u00a0Procedure: Monkeypox\u00a0Virus\u00a0Generic\u00a0Real\u2010time PCR Test","author":"CDC.\u00a0","year":"2022"},{"key":"e_1_2_9_11_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jviromet.2010.07.012"},{"key":"e_1_2_9_12_1","doi-asserted-by":"publisher","DOI":"10.2807\/1560-7917.ES.2022.27.28.2200503"},{"key":"e_1_2_9_13_1","article-title":"Detection of monkeypox viral DNA in a routine wastewater monitoring program","author":"Wolfe MK","year":"2022","journal-title":"medRxiv"},{"key":"e_1_2_9_14_1","article-title":"An accurate and rapid Real\u2010time PCR approach for human Monkeypox virus diagnosis","author":"Wawina\u2010Bokalanga T","year":"2022","journal-title":"medRxiv"},{"key":"e_1_2_9_15_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jviromet.2016.07.024"},{"key":"e_1_2_9_16_1","doi-asserted-by":"publisher","DOI":"10.1177\/153567601502000405"},{"key":"e_1_2_9_17_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.mcp.2009.10.008"},{"key":"e_1_2_9_18_1","unstructured":"CDC. 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