{"id":1138,"date":"2013-01-16T13:39:06","date_gmt":"2013-01-16T12:39:06","guid":{"rendered":"http:\/\/sppadbase.ipp.cnr.it\/?p=1138"},"modified":"2013-01-16T13:46:22","modified_gmt":"2013-01-16T12:46:22","slug":"development-of-a-loop-mediated-isothermal-amplification-assay-for-detection-of-phytophthora-sojae","status":"publish","type":"post","link":"https:\/\/sppadbase.ipsp.cnr.it\/?p=1138","title":{"rendered":"Development of a loop-mediated isothermal amplification assay for detection of Phytophthora sojae"},"content":{"rendered":"<div class=\"textimage\">\n<p class=\"textimage\"><a href=\"http:\/\/sppadbase.ipp.cnr.it\/?attachment_id=1331\" rel=\"attachment wp-att-1331\"><img decoding=\"async\" loading=\"lazy\" src=\"http:\/\/sppadbase.ipp.cnr.it\/wordpress\/wp-content\/uploads\/2013\/01\/fml2619-fig-0001-295x300.png\" alt=\"fml2619-fig-0001\" width=\"295\" height=\"300\" class=\"alignnone size-medium wp-image-1331\" srcset=\"https:\/\/sppadbase.ipsp.cnr.it\/wp-content\/uploads\/2013\/01\/fml2619-fig-0001-295x300.png 295w, https:\/\/sppadbase.ipsp.cnr.it\/wp-content\/uploads\/2013\/01\/fml2619-fig-0001-147x150.png 147w, https:\/\/sppadbase.ipsp.cnr.it\/wp-content\/uploads\/2013\/01\/fml2619-fig-0001-400x405.png 400w, https:\/\/sppadbase.ipsp.cnr.it\/wp-content\/uploads\/2013\/01\/fml2619-fig-0001.png 422w\" sizes=\"(max-width: 295px) 100vw, 295px\" \/><\/a><em>Phytophthora sojae<\/em> is a devastating pathogen that causes soybean <em>Phytophthora<\/em> root rot. This study reports the development of a loop-mediated isothermal amplification (LAMP) assay targeting the <em>A3aPro<\/em> element for visual detection of <em>P.\u00a0sojae<\/em>. The <em>A3aPro<\/em>-LAMP assay efficiently amplified the target element in &lt;\u00a080\u00a0min at 64\u00a0\u00b0C and was evaluated for specificity and sensitivity. The specificity was evaluated against <em>P.\u00a0sojae<\/em>, <em>Phytophthora<\/em> spp., <em>Pythium<\/em> spp., and true fungi isolates. Magnesium pyrophosphate resulting from the LAMP of P.\u00a0sojae could be detected by real-time measurement of turbidity. <em>Phytophthora sojae<\/em> DNA products were visualized as a ladder-like banding pattern on 2% gel electrophoresis. A positive colour (sky blue) was only observed in the presence of <em>P.\u00a0sojae<\/em> with the addition of hydroxynaphthol blue prior to amplification, whereas none of other isolates showed a colour change. The detection limit of the <em>A3aPro<\/em>-specific LAMP assay for<em> P.\u00a0sojae<\/em> was 10\u00a0pg\u00a0\u03bcL\u22121 of genomic DNA per reaction. The assay also detected <em>P.\u00a0sojae<\/em> from diseased soybean tissues and residues. These results suggest that the <em>A3aPro<\/em>-LAMP assay reported here can be used for the visual detection of <em>P.\u00a0sojae<\/em> in plants and production fields.<\/p>\n<div class=\"clearer\"><\/div>\n<blockquote><p>The paper reports other primer pairs for an alternative PCR based detection technology, the loop-mediated isothermal amplification (LAMP) of DNA. <strong><em>The current database architecture does not support LAMP primers<\/em><\/strong>. Only the forward and backward outer primers will be loaded. Users are asked to check the original paper for primer sequences and details.<\/p><\/blockquote>\n<p><strong>Ting-Ting Dai, Chen-Chen Lu, Jing Lu, SuoMeng Dong, WenWu Ye, YuanChao Wang, XiaoBo Zheng<\/strong><br \/>\nFEMS Microbiology Letters<br \/>\nVolume 334, Issue 1, pages 27\u201334, September 2012<\/p>\n<p>DOI: 10.1111\/j.1574-6968.2012.02619.x<\/p>\n<p>via <a href='http:\/\/onlinelibrary.wiley.com\/doi\/10.1111\/j.1574-6968.2012.02619.x\/full'>Development of a loop-mediated isothermal amplification assay for detection of Phytophthora sojae &#8211; Dai &#8211; 2012 &#8211; FEMS Microbiology Letters &#8211; Wiley Online Library<\/a>.<\/p>\n","protected":false},"excerpt":{"rendered":"\n<p class=\"textimage\">Phytophthora sojae is a devastating pathogen that causes soybean Phytophthora root rot. This study reports the development of a loop-mediated isothermal amplification (LAMP) assay targeting the A3aPro element for visual detection of P. sojae. The A3aPro-LAMP assay efficiently amplified the target element in &lt; 80 min at 64 \u00b0C and was evaluated for <span style=\"color:#777\"> . . . &rarr; Read More: <a href=\"https:\/\/sppadbase.ipsp.cnr.it\/?p=1138\">Development of a loop-mediated isothermal amplification assay for detection of Phytophthora sojae<\/a><\/span><\/p>\n","protected":false},"author":1,"featured_media":0,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":[],"categories":[15,81,3,11,27,101],"tags":[23,16,116,58,119,161,19,59],"_links":{"self":[{"href":"https:\/\/sppadbase.ipsp.cnr.it\/index.php?rest_route=\/wp\/v2\/posts\/1138"}],"collection":[{"href":"https:\/\/sppadbase.ipsp.cnr.it\/index.php?rest_route=\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/sppadbase.ipsp.cnr.it\/index.php?rest_route=\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/sppadbase.ipsp.cnr.it\/index.php?rest_route=\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/sppadbase.ipsp.cnr.it\/index.php?rest_route=%2Fwp%2Fv2%2Fcomments&post=1138"}],"version-history":[{"count":13,"href":"https:\/\/sppadbase.ipsp.cnr.it\/index.php?rest_route=\/wp\/v2\/posts\/1138\/revisions"}],"predecessor-version":[{"id":1338,"href":"https:\/\/sppadbase.ipsp.cnr.it\/index.php?rest_route=\/wp\/v2\/posts\/1138\/revisions\/1338"}],"wp:attachment":[{"href":"https:\/\/sppadbase.ipsp.cnr.it\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=1138"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/sppadbase.ipsp.cnr.it\/index.php?rest_route=%2Fwp%2Fv2%2Fcategories&post=1138"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/sppadbase.ipsp.cnr.it\/index.php?rest_route=%2Fwp%2Fv2%2Ftags&post=1138"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}