{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,19]],"date-time":"2026-06-19T04:41:43Z","timestamp":1781844103087,"version":"3.54.5"},"reference-count":23,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2022,2,28]],"date-time":"2022-02-28T00:00:00Z","timestamp":1646006400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003993","name":"Ministry of Agriculture, Forestry and Fisheries of Japan","doi-asserted-by":"publisher","award":["JPJ008617. 20319736"],"award-info":[{"award-number":["JPJ008617. 20319736"]}],"id":[{"id":"10.13039\/501100003993","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>African swine fever (ASF) and classical swine fever (CSF) are contagious swine diseases that are clinically indistinguishable from each other; hence, reliable test methods for accurate diagnosis and differentiation are highly demanded. By employing a buffer system suitable for crude extraction of nucleic acids together with an impurity-tolerant enzyme, we established a multiplex assay of real-time reverse-transcription polymerase chain reaction (rRT-PCR) for simultaneous detection of ASF virus (ASFV), CSF virus (CSFV) and swine internal control derived genes in a sample without the need for prior purification of viral nucleic acids. We applied this method to test serum and tissue samples of infected pigs and wild boars and compared the statistical sensitivities and specificities with those of standard molecular diagnostic methods. When a serum was used as a test material, the newly established assay showed 94.4% sensitivity for both and 97.9 and 91.9% specificity for ASFV and CSFV detection, respectively. In contrast, the results were 100% identical with those obtained by the standard methods when a crude tissue homogenate was used as a test material. The present data indicate that this new assay offers a practical, quick, and reliable technique for differential diagnosis of ASF and CSF where geographical occurrences are increasingly overlapping.<\/jats:p>","DOI":"10.3390\/v14030498","type":"journal-article","created":{"date-parts":[[2022,2,28]],"date-time":"2022-02-28T20:11:57Z","timestamp":1646079117000},"page":"498","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":25,"title":["Establishment of a Direct PCR Assay for Simultaneous Differential Diagnosis of African Swine Fever and Classical Swine Fever Using Crude Tissue Samples"],"prefix":"10.3390","volume":"14","author":[{"given":"Tatsuya","family":"Nishi","sequence":"first","affiliation":[{"name":"Exotic Disease Research Station, National Institute of Animal Health, National Agriculture and Food Research Organization, 6-20-1 Josui-honcho, Kodaira 187-0022, Tokyo, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kota","family":"Okadera","sequence":"additional","affiliation":[{"name":"Exotic Disease Research Station, National Institute of Animal Health, National Agriculture and Food Research Organization, 6-20-1 Josui-honcho, Kodaira 187-0022, Tokyo, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Katsuhiko","family":"Fukai","sequence":"additional","affiliation":[{"name":"Exotic Disease Research Station, National Institute of Animal Health, National Agriculture and Food Research Organization, 6-20-1 Josui-honcho, Kodaira 187-0022, Tokyo, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Miwa","family":"Yoshizaki","sequence":"additional","affiliation":[{"name":"Takara Bio Inc., Nojihigashi 7-4-38, Kusatsu 525-0058, Shiga, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ai","family":"Nakasuji","sequence":"additional","affiliation":[{"name":"Takara Bio Inc., Nojihigashi 7-4-38, Kusatsu 525-0058, Shiga, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Syuji","family":"Yoneyama","sequence":"additional","affiliation":[{"name":"Tochigi Prefectural Central District Animal Hygiene Service Centre, Hiraideko-gyo-danchi 6-8, Utsunomiya 321-0905, Tchigi, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Takehiro","family":"Kokuho","sequence":"additional","affiliation":[{"name":"Exotic Disease Research Station, National Institute of Animal Health, National Agriculture and Food Research Organization, 6-20-1 Josui-honcho, Kodaira 187-0022, Tokyo, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,2,28]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"122","DOI":"10.1016\/j.virusres.2012.10.026","article-title":"Pathogenesis of African Swine Fever in Domestic Pigs and European Wild Boar","volume":"173","author":"Blome","year":"2013","journal-title":"Virus Res."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Blome, S., Staubach, C., Henke, J., Carlson, J., and Beer, M. 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