{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,4]],"date-time":"2026-05-04T21:41:06Z","timestamp":1777930866415,"version":"3.51.4"},"reference-count":20,"publisher":"SAGE Publications","issue":"3","license":[{"start":{"date-parts":[[2023,3,12]],"date-time":"2023-03-12T00:00:00Z","timestamp":1678579200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.sagepub.com\/page\/policies\/text-and-data-mining-license"}],"funder":[{"name":"AMP Core of the Center of Emerging and Zoonotic Infectious Diseases (CEZID) from National Institute of General Medical Sciences","award":["P20GM130448"],"award-info":[{"award-number":["P20GM130448"]}]},{"name":"the Department of Homeland Security for the Center of Excellence for Emerging and Zoonotic Diseases","award":["DHS 2010-ST-061-AG0001"],"award-info":[{"award-number":["DHS 2010-ST-061-AG0001"]}]}],"content-domain":{"domain":["journals.sagepub.com"],"crossmark-restriction":true},"short-container-title":["J VET Diagn Invest"],"published-print":{"date-parts":[[2023,5]]},"abstract":"<jats:p>African swine fever (ASF) causes fatal disease in pigs and is an escalating threat to the global swine industry. ASF has re-emerged from Africa as a transcontinental epidemic spreading through the Caucasus into Europe, Russia, China, numerous Asian countries, and the Caribbean. ASF virus (ASFV) is a U.S. select agent requiring handling in high-containment biosafety level 3 (BSL-3) laboratories for pathogen work. Formalin-fixation eliminates infectivity and preserves the genome, providing noninfectious specimens for BSL-2 work. Recovery of DNA from formalin-fixed, paraffin-embedded tissue (FFPET) is challenging and cumbersome. A reliable and easy-to-perform method for DNA recovery from FFPET would facilitate surveillance. To meet this objective, we developed a high-throughput protocol for the recovery of ASFV DNA from FFPET. Deparaffinization, tissue lysis, and reversal of cross-linking were performed in a single tube, followed by DNA purification via automated magnetic bead extraction. Quantitative PCR (qPCR) detection was used to determine the copy number of the B646L gene that encodes for the ASFV p72 protein in tissues (5 pigs, 4 tissues) from pigs with lesions consistent with acute ASF. Copy numbers obtained from FFPET were within one log of copy numbers obtained from fresh tissue, thus enabling ASF qPCR surveillance from formalin-inactivated and preserved tissues at BSL-2 at diagnostic sensitivity similar to fresh tissues tested at BSL-3.<\/jats:p>","DOI":"10.1177\/10406387231158534","type":"journal-article","created":{"date-parts":[[2023,3,13]],"date-time":"2023-03-13T02:28:35Z","timestamp":1678674515000},"page":"284-288","update-policy":"https:\/\/doi.org\/10.1177\/sage-journals-update-policy","source":"Crossref","is-referenced-by-count":0,"title":["Development of a sensitive, high-throughput extraction protocol for qPCR detection of African swine fever virus in formalin-fixed, paraffin-embedded tissues"],"prefix":"10.1177","volume":"35","author":[{"given":"Kinga","family":"Urbaniak","sequence":"first","affiliation":[{"name":"Department of Diagnostic Medicine\/Pathobiology, College of Veterinary Medicine, Kansas State University, Manhattan, KS, USA"},{"name":"Center of Excellence for Emerging and Zoonotic Animal Diseases, College of Veterinary Medicine, Kansas State University, Manhattan, KS, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"David A.","family":"Meekins","sequence":"additional","affiliation":[{"name":"Department of Diagnostic Medicine\/Pathobiology, College of Veterinary Medicine, Kansas State University, Manhattan, KS, USA"},{"name":"Center of Excellence for Emerging and Zoonotic Animal Diseases, College of Veterinary Medicine, Kansas State University, Manhattan, KS, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5711-3936","authenticated-orcid":false,"given":"A. Sally","family":"Davis","sequence":"additional","affiliation":[{"name":"Department of Diagnostic Medicine\/Pathobiology, College of Veterinary Medicine, Kansas State University, Manhattan, KS, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Juergen A.","family":"Richt","sequence":"additional","affiliation":[{"name":"Department of Diagnostic Medicine\/Pathobiology, College of Veterinary Medicine, Kansas State University, Manhattan, KS, USA"},{"name":"Center of Excellence for Emerging and Zoonotic Animal Diseases, College of Veterinary Medicine, Kansas State University, Manhattan, KS, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5644-022X","authenticated-orcid":false,"given":"Jessie D.","family":"Trujillo","sequence":"additional","affiliation":[{"name":"Department of Diagnostic Medicine\/Pathobiology, College of Veterinary Medicine, Kansas State University, Manhattan, KS, USA"},{"name":"Center of Excellence for Emerging and Zoonotic Animal Diseases, College of Veterinary Medicine, Kansas State University, Manhattan, KS, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"179","published-online":{"date-parts":[[2023,3,12]]},"reference":[{"key":"bibr1-10406387231158534","doi-asserted-by":"publisher","DOI":"10.1016\/S0378-1135(98)00187-4"},{"key":"bibr2-10406387231158534","first-page":"161","volume":"62","author":"Cisek AA","year":"2016","journal-title":"Ann Parasitol"},{"key":"bibr3-10406387231158534","doi-asserted-by":"publisher","DOI":"10.1016\/j.vetmic.2012.06.025"},{"key":"bibr4-10406387231158534","doi-asserted-by":"crossref","unstructured":"Dixon LK, et al. 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