{"id":2506,"date":"2019-11-29T18:39:28","date_gmt":"2019-11-30T02:39:28","guid":{"rendered":"https:\/\/labs.wsu.edu\/tanaka-lab\/?p=2506"},"modified":"2022-02-28T09:47:46","modified_gmt":"2022-02-28T17:47:46","slug":"rpa-powdery-scab-pmtv","status":"publish","type":"post","link":"https:\/\/labs.wsu.edu\/tanaka-lab\/2019\/11\/29\/rpa-powdery-scab-pmtv\/","title":{"rendered":"RPA assays developed for the powdery scab pathogen and PMTV published in AJPR"},"content":{"rendered":"<p><strong>November 29, 2019<\/strong> \u2013 Our article entitled \u201cRecombinase Polymerase Amplification (RPA) for the Rapid Isothermal Detection of <em>Spongospora subterrane<\/em>a f. sp. <em>subterranea<\/em> and Potato Mop-Top Virus\u201d was published in <em>American Journal of Potato Research<\/em>.\u00a0<!--more--><\/p>\n<p><strong>How to cite:<br \/>\n<\/strong>DeShields JB, Moroz N, Braley LE, Mora-Romero GA, Tanaka K (2019) Recombinase polymerase amplification (RPA) for the rapid isothermal detection of <em>Spongospora subterranea<\/em> f. sp. <em>subterranea<\/em> and potato mop-top virus. <em>American Journal of Potato Research<\/em> 96: 617-624 <a href=\"https:\/\/doi.org\/10.1007\/s12230-019-09750-7\">doi:10.1007\/s12230-019-09750-7<\/a><\/p>\n<p><strong>Abstract:<\/strong><br \/>\nThe amplification of specific nucleic acid sequences with high specificity and sensitivity is an essential technique for pathogen detection. Recombinase polymerase amplification (RPA) is a rapid isothermal amplification method. Here, we demonstrate the end-point and real-time detection of <em>Spongospora subterranea<\/em> f. sp. <em>subterranea<\/em> (Sss) using RPA and potato mop-top virus (PMTV) using reverse transcription (RT)-RPA. Oligonucleotide primers were designed for amplification that target the internal transcribed spacer 1 (ITS1) region and the coat protein readthrough (CP-RT) domain for the detection of Sss and PMTV, respectively. Our data showed that real-time RPA can detect 100 of Sss sporosori per gram of soil, while real-time RT-RPA could detect in ~1 ng total RNA of the PMTV-infected tuber. For Sss detection, the R2 value for real-time RPA and real-time PCR was 98.0% by linear regression analysis in the concentration range of 100\u201334,000 sporosori per gram of soil. The developed RPA assay here may provide a useful alternative tool for the rapid, simple and reliable detection of Sss and PMTV in diagnostic laboratories and in-field testing.<\/p>\n<p>&nbsp;<\/p>\n","protected":false},"excerpt":{"rendered":"<p><strong>November 29, 2019<\/strong> \u2013 Our article entitled \u201cRecombinase Polymerase Amplification (RPA) for the Rapid Isothermal Detection of <em>Spongospora subterrane<\/em>a f. sp. <em>subterranea<\/em> and Potato Mop-Top Virus\u201d was published in <em>American Journal of Potato Research<\/em>.\u00a0<\/p>\n","protected":false},"author":2755,"featured_media":2439,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":[],"categories":[],"tags":[],"wsuwp_university_location":[],"wsuwp_university_org":[],"_links":{"self":[{"href":"https:\/\/labs.wsu.edu\/tanaka-lab\/wp-json\/wp\/v2\/posts\/2506"}],"collection":[{"href":"https:\/\/labs.wsu.edu\/tanaka-lab\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/labs.wsu.edu\/tanaka-lab\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/labs.wsu.edu\/tanaka-lab\/wp-json\/wp\/v2\/users\/2755"}],"replies":[{"embeddable":true,"href":"https:\/\/labs.wsu.edu\/tanaka-lab\/wp-json\/wp\/v2\/comments?post=2506"}],"version-history":[{"count":8,"href":"https:\/\/labs.wsu.edu\/tanaka-lab\/wp-json\/wp\/v2\/posts\/2506\/revisions"}],"predecessor-version":[{"id":2908,"href":"https:\/\/labs.wsu.edu\/tanaka-lab\/wp-json\/wp\/v2\/posts\/2506\/revisions\/2908"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/labs.wsu.edu\/tanaka-lab\/wp-json\/wp\/v2\/media\/2439"}],"wp:attachment":[{"href":"https:\/\/labs.wsu.edu\/tanaka-lab\/wp-json\/wp\/v2\/media?parent=2506"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/labs.wsu.edu\/tanaka-lab\/wp-json\/wp\/v2\/categories?post=2506"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/labs.wsu.edu\/tanaka-lab\/wp-json\/wp\/v2\/tags?post=2506"},{"taxonomy":"wsuwp_university_location","embeddable":true,"href":"https:\/\/labs.wsu.edu\/tanaka-lab\/wp-json\/wp\/v2\/wsuwp_university_location?post=2506"},{"taxonomy":"wsuwp_university_org","embeddable":true,"href":"https:\/\/labs.wsu.edu\/tanaka-lab\/wp-json\/wp\/v2\/wsuwp_university_org?post=2506"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}