{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,12]],"date-time":"2026-08-12T15:53:39Z","timestamp":1786550019444,"version":"build-2736575974"},"reference-count":23,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2021,2,25]],"date-time":"2021-02-25T00:00:00Z","timestamp":1614211200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Indical Bioscience GmbH","award":["Ri-0739"],"award-info":[{"award-number":["Ri-0739"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>African swine fever (ASF) is a contagious viral hemorrhagic disease of domestic pigs and wild boars. The disease is notifiable to the World Organisation for Animal Health (OIE) and is responsible for high mortality and serious economic losses. PCR and real-time PCR (qPCR) are the OIE-recommended standard methods for the direct detection of African swine fever virus (ASFV) DNA. The aim of our work was the simplification and standardization of the molecular diagnostic workflow in the lab. For validation of this \u201ceasy lab\u201d workflow, different sample materials from animal trials were collected and analyzed (EDTA blood, serum, oral swabs, chewing ropes, and tissue samples) to identify the optimal sample material for diagnostics in live animals. Based on our data, the EDTA blood samples or bloody tissue samples represent the best specimens for ASFV detection in the early and late phases of infection. The application of prefilled ready-to-use reagents for nucleic acid extraction or the use of a Tissue Lysis Reagent (TLR) delivers simple and reliable alternatives for the release of the ASFV nucleic acids. For the qPCR detection of ASFV, different published and commercial kits were compared. Here, a lyophilized commercial kit shows the best results mainly based on the increased template input. The good results of the \u201ceasy lab\u201d strategy could be confirmed by the ASFV detection in field samples from wild boars collected from the 2020 ASFV outbreak in Germany. Appropriate internal control systems for extraction and PCR are key features of the \u201ceasy lab\u201d concept and reduce the risk of false-negative and false-positive results. In addition, the use of easy-to-handle machines and software reduces training efforts and the misinterpretation of results. The PCR diagnostics based on the \u201ceasy lab\u201d strategy can realize a high sensitivity and specificity comparable to the standard PCR methods and should be especially usable for labs with limited experiences and resources.<\/jats:p>","DOI":"10.3390\/ijms22052307","type":"journal-article","created":{"date-parts":[[2021,2,26]],"date-time":"2021-02-26T02:08:17Z","timestamp":1614305297000},"page":"2307","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":16,"title":["Swift and Reliable \u201cEasy Lab\u201d Methods for the Sensitive Molecular Detection of African Swine Fever Virus"],"prefix":"10.3390","volume":"22","author":[{"given":"Ahmed","family":"Elnagar","sequence":"first","affiliation":[{"name":"Friedrich-Loeffler-Institut, Institute of Diagnostic Virology, 17493 Greifswald-Insel Riems, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3502-4240","authenticated-orcid":false,"given":"Jutta","family":"Pikalo","sequence":"additional","affiliation":[{"name":"Friedrich-Loeffler-Institut, Institute of Diagnostic Virology, 17493 Greifswald-Insel Riems, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Martin","family":"Beer","sequence":"additional","affiliation":[{"name":"Friedrich-Loeffler-Institut, Institute of Diagnostic Virology, 17493 Greifswald-Insel Riems, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5465-5609","authenticated-orcid":false,"given":"Sandra","family":"Blome","sequence":"additional","affiliation":[{"name":"Friedrich-Loeffler-Institut, Institute of Diagnostic Virology, 17493 Greifswald-Insel Riems, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5358-6445","authenticated-orcid":false,"given":"Bernd","family":"Hoffmann","sequence":"additional","affiliation":[{"name":"Friedrich-Loeffler-Institut, Institute of Diagnostic Virology, 17493 Greifswald-Insel Riems, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,2,25]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"2683","DOI":"10.1098\/rstb.2009.0098","article-title":"African swine fever: How can global spread be prevented?","volume":"364","author":"Costard","year":"2009","journal-title":"Philos. 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