{"id":9905,"date":"2016-09-30T15:07:36","date_gmt":"2016-09-30T06:07:36","guid":{"rendered":"http:\/\/cell.brc.riken.jp\/ja\/?page_id=9905"},"modified":"2026-02-16T15:02:28","modified_gmt":"2026-02-16T06:02:28","slug":"ebv","status":"publish","type":"page","link":"http:\/\/cell.brc.riken.jp\/ja\/quality\/virus\/ebv","title":{"rendered":"EBV\u691c\u67fb"},"content":{"rendered":"<p>EB\u30a6\u30a4\u30eb\u30b9\uff08Epstein-Barr virus; EBV\uff09\u306f\u3001\u30d8\u30eb\u30da\u30b9\u30a6\u30a3\u30eb\u30b9\u79d1\u306b\u5c5e\u3059\u308b\u75c5\u539f\u6027\u30a6\u30a4\u30eb\u30b9\u306e\u4e00\u7a2e\u3067\u3042\u308a\u3001\u553e\u6db2\u3092\u4ecb\u3057\u3066\u611f\u67d3\u3092\u5e83\u3052\u3001\u4e3b\u306bB\u7d30\u80de\u306b\u611f\u67d3\u3057\u307e\u3059\u3002EBV\u306f95%\u4ee5\u4e0a\u306e\u5065\u5e38\u6210\u4eba\u304c\u611f\u67d3\u3057\u3001\u305d\u306e\u591a\u304f\u304c\u7d42\u751f\u5171\u751f\u7684\u306b\u4fdd\u6301\u3055\u308c\u308b\u8ab0\u3082\u304c\u611f\u67d3\u3057\u3066\u3044\u308b\u30a6\u30a4\u30eb\u30b9\u3067\u3042\u308b\u4e00\u65b9\u3001\u767a\u75c7\u306f\u7a00\u3067\u306f\u3042\u308a\u307e\u3059\u304c\u3001\u30d0\u30fc\u30ad\u30c3\u30c8\u30ea\u30f3\u30d1\u816b\u306a\u3069\u4e00\u90e8\u306e\u60aa\u6027\u30ea\u30f3\u30d1\u816b\u3084\u4e0a\u54bd\u982d\u304c\u3093\u306e\u767a\u751f\u3068\u95a2\u9023\u304c\u3042\u308b\u3053\u3068\u304c\u660e\u3089\u304b\u306b\u306a\u3063\u3066\u3044\u307e\u3059\u3002<\/p>\n<p>\u5f53\u5ba4\u3067\u306f\u3001\u30d2\u30c8\u8840\u6db2\u7531\u6765\u7d30\u80de\u682a\uff08EBV\u5f62\u8cea\u8ee2\u63dbB\u7d30\u80de\u682a\u3092\u9664\u304f\uff09\u3084EBV\u611f\u67d3\u304c\u764c\u767a\u75c7\u306e\u539f\u56e0\u3068\u306e\u5831\u544a\u304c\u3042\u308b\u30d2\u30c8\u764c\u7d30\u80de\u682a\uff08\u53e3\u8154\u764c\u3001\u98df\u9053\u764c\u3001\u80c3\u764c\uff09\u306b\u3064\u3044\u3066\u3001\u6f5c\u4f0f\u611f\u67d3\u95a2\u9023\u907a\u4f1d\u5b50\u3067\u3042\u308bEBNA1<sup>*1<\/sup>\u307e\u305f\u306f\u6eb6\u89e3\u611f\u67d3\u95a2\u9023\u907a\u4f1d\u5b50\u3067\u3042\u308bBALF5<sup>*2<\/sup>\u3092\u6307\u6a19\u3068\u3057\u3001\u7d30\u80de\u306b\u611f\u67d3\u3057\u305fEBV\u30b2\u30ce\u30e0\u3092qPCR\u6cd5\u306b\u3088\u308a\u691c\u51fa\u3057\u3066\u3044\u307e\u3059\u3002<br \/>\n<sup>*1<\/sup> EBNA1: Epstein-Barr Nuclear Antigen 1<br \/>\n<sup>*2<\/sup> BALF5: the BamHI-A fragment containing the fifth leftward open reading frame<\/p>\n<p><strong>1. \u6e96\u5099\u3059\u308b\u3082\u306e<\/strong><br \/>\n(1) \u691c\u4f53\uff1a\u7d30\u80de\u304b\u3089\u62bd\u51fa\u3057\u305f\u30b2\u30ce\u30e0DNA\u6eb6\u6db2<br \/>\n(2) \u6a5f\u5668\uff1a Applied Biosystems QuantStudio3 Real-Time PCR System<br \/>\n(3) \u30bd\u30d5\u30c8\u30a6\u30a7\u30a2\uff1aQuantStudio Design &#038; Analysis Software\u3001ThermoFisher SCIENTIFIC<br \/>\n(4) PCR \u8a66\u85ac\uff1aApplied Biosystems PowerUp SYBR Green Master Mix (Cat No. A25777\uff09<br \/>\n(5) PCR\u30d7\u30e9\u30a4\u30de\u30fc<\/p>\n<ul>\n<li>EBNA1\u30d7\u30e9\u30a4\u30de\u30fc<\/li>\n<p>\u3000EBNA1-F : <tt>5\u2019-ATCAGGGCCAAGACATAGAGATG-3\u2019<\/tt><br \/>\n\u3000EBNA1-R : <tt>5\u2019-GCCAATGCAACTTGGACGTT-3\u2019<\/tt>\n<\/ul>\n<ul>\n<li>BALF5\u30d7\u30e9\u30a4\u30de\u30fc<\/li>\n<p>\u3000BALF5-F : <tt>5\u2019-CGGAAGCCCTCTGGACTTC\u20133\u2019<\/tt><br \/>\n\u3000BALF5-R : <tt>5\u2019-CCCTGTTTATCCGATGGAATG\u20133\u2019<\/tt>\n<\/ul>\n<p>(6) \u30b9\u30bf\u30f3\u30c0\u30fc\u30c9\uff1a\u305d\u308c\u305e\u308c\u306e\u30bf\u30fc\u30b2\u30c3\u30c8\u907a\u4f1d\u5b50\u306b\u3064\u3044\u3066\u3001\u5408\u6210DNA\u3092\u5229\u7528\u3059\u308b\u3002<\/p>\n<p><strong>2. \u65b9\u6cd5<\/strong><br \/>\n(1) \u30b9\u30bf\u30f3\u30c0\u30fc\u30c9\u306e\u8abf\u88fd\uff1a\u305d\u308c\u305e\u308c\u306e\u30bf\u30fc\u30b2\u30c3\u30c8\u907a\u4f1d\u5b50\u306b\u3064\u3044\u3066\u4ee5\u4e0b\u306e\u30b3\u30d4\u30fc\u6570\u3092\u8abf\u88fd\u3059\u308b\u3002<\/p>\n<ul>\n<li>EBNA1\uff1b1&#215;10<sup>6<\/sup>\u30011&#215;10<sup>5<\/sup>\u30011&#215;10<sup>4<\/sup>\u30011&#215;10<sup>3<\/sup>\u30011&#215;10<sup>2<\/sup>\u30011&#215;10<sup>1<\/sup> (copies)<\/li>\n<li>BALF5\uff1b2.5&#215;10<sup>6<\/sup>\u30012.5&#215;10<sup>5<\/sup>\u30012.5&#215;10<sup>4<\/sup>\u30012.5&#215;10<sup>3<\/sup>\u30012.5&#215;10<sup>2<\/sup>\u30012.5&#215;10<sup>1<\/sup> (copies)<\/li>\n<\/ul>\n<p>(2) \u691c\u4f53\u306e\u8abf\u88fd\uff1a2.5 ng\/uL \u30b2\u30ce\u30e0DNA\u6eb6\u6db2\u3092\u8abf\u88fd<br \/>\n(3) PCR\u30de\u30b9\u30bf\u30fc\u30df\u30c3\u30af\u30b9\u306e\u8abf\u88fd<\/p>\n<table style=\"margin-left:20px;\">\n<tbody>\n<tr>\n<td id=\"t1\">DDW<\/td>\n<td id=\"t1\" style=\"text-align: right;\">4 uL<\/td>\n<\/tr>\n<tr>\n<td id=\"t1\">\u30d7\u30e9\u30a4\u30de\u30fcFW (10 pmol\/\u00b5l)<\/td>\n<td id=\"t1\" style=\"text-align: right;\">1 uL<\/td>\n<\/tr>\n<tr>\n<td id=\"t1\">\u30d7\u30e9\u30a4\u30de\u30fcRV (10 pmol\/\u00b5l)<\/td>\n<td id=\"t1\" style=\"text-align: right;\">1 uL<\/td>\n<\/tr>\n<tr>\n<td id=\"t1\">PowerUp SYBR Green Master Mix (2x)<\/td>\n<td id=\"t1\" style=\"text-align: right;\">10 uL<\/td>\n<\/tr>\n<tr>\n<td colspan=\"2\" id=\"t1\">\n<hr \/>\n<\/td>\n<\/tr>\n<tr>\n<td id=\"t1\">Total<\/td>\n<td id=\"t1\" style=\"text-align: right;\">16 uL \/Sample<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<p>(4) PCR\u752896 well plate\u306b\u30de\u30b9\u30bf\u30fc\u30df\u30c3\u30af\u30b9\u309216 uL\u305a\u3064\u5206\u6ce8\u3059\u308b\u3002<br \/>\n(5) (2)\u3067\u7528\u610f\u3057\u305f\u691c\u4f53\u30924 uL\u305a\u3064\u52a0\u3048\u308b\u3002<br \/>\n(6) PCR program<br \/>\n\u3000\u3000UDG\u6d3b\u6027\u5316\uff1a50\u2103 2\u5206 (\u30db\u30fc\u30eb\u30c9\uff09<br \/>\n\u3000\u3000Dual-Lock\u2019DNA \u30dd\u30ea\u30e1\u30e9\u30fc\u30bc\uff1a95\u2103 2\u5206 (\u30db\u30fc\u30eb\u30c9\uff09<br \/>\n\u3000\u3000\u53cd\u5fdc\u30b5\u30a4\u30af\u30eb(\u5909\u6027\u2192\u30a2\u30cb\u30fc\u30ea\u30f3\u30b0\/\u4f38\u9577)\uff1a95\u2103 15\u79d2 \u2192 60\u2103 1\u5206 <sup>*<\/sup> \uff0840\u30b5\u30a4\u30af\u30eb\uff09<br \/>\n\u3000\u3000\u878d\u89e3\u66f2\u7dda\u89e3\u6790\uff1a95\u2103 15\u79d2 \u2192 60\u2103 1\u5206 \u2192 95\u2103 15\u79d2 <sup>*<\/sup> (\uff11\u30b5\u30a4\u30af\u30eb\uff09<br \/>\n\u3000\u3000\u3000\u3000\u3000<sup>*<\/sup> \u86cd\u5149\u30c7\u30fc\u30bf\u56de\u53ce\u30dd\u30a4\u30f3\u30c8<br \/>\n(7) \u7d50\u679c\u89e3\u6790<\/p>\n<p><strong>3. \u5224\u5b9a<\/strong><br \/>\n\u691c\u67fb\u306e\u6709\u52b9\u6027 (\u30b9\u30bf\u30f3\u30c0\u30fc\u30c9\u306eEfficiency\u3001R<sup>2<\/sup>\u3001Melt Curve\u306eTM\u5024\u306a\u3069\u304c\u6709\u52b9\u6761\u4ef6\u5185\u3067\u3042\u308b\u3053\u3068)\u306e\u78ba\u8a8d\u5f8c\u3001\u5224\u5b9a\u3092\u884c\u3046\u3002<\/p>\n<p><strong>4. \u53c2\u8003\u6587\u732e<\/strong><br \/>\n<u>PCR\u306b\u95a2\u3059\u308b\u53c2\u8003\u6587\u732e<\/u><br \/>\nQuantStudio 3\/QuantStudio 5 \u30ea\u30a2\u30eb\u30bf\u30a4\u30e0 PCR \u30b7\u30b9\u30c6\u30e0 \u7c21\u6613\u64cd\u4f5c\u30ac\u30a4\u30c9<\/p>\n<p><u>\u691c\u51fa\u9650\u754c\u5024\u6c7a\u5b9a\u691c\u67fb\u306b\u95a2\u3059\u308b\u53c2\u8003\u6587\u732e<\/u><br \/>\n\u8840\u6db2\u88fd\u5264\u306e\u30a6\u30a4\u30eb\u30b9\u306b\u5bfe\u3059\u308b\u5b89\u5168\u6027\u78ba\u4fdd\u3092\u76ee\u7684\u3068\u3057\u305f\u6838\u9178\u5897\u5e45\u691c\u67fb(NAT)\u306e\u5b9f\u65bd\u306b\u95a2\u3059\u308b\u30ac\u30a4\u30c9\u30e9\u30a4\u30f3 (\u539a\u751f\u52b4\u50cd\u7701)<\/p>\n<p><u>Primer\u3084\u30b9\u30bf\u30f3\u30c0\u30fc\u30c9\u306b\u95a2\u3059\u308b\u53c2\u8003\u6587\u732e<\/u><br \/>\nGeneration of two human induced pluripotent stem cell lines derived from two juvenile nephronophthisis patients with NPHP1 deletion. <em>Stem Cell Res. <\/em>2020 May; 45:101815. <a href=\"https:\/\/pubmed.ncbi.nlm.nih.gov\/32361464\/\">PMID: 32361464.<\/a><\/p>\n<p>Screening for 15 pathogenic viruses in human cell lines registered at the JCRB Cell Bank: characterization of <em>in vitro<\/em> human cells by viral infection. <em>R Soc Open Sci.<\/em> 2018 May 2;5(5):172472. <a href=\"https:\/\/royalsocietypublishing.org\/doi\/10.1098\/rsos.172472\">PMID: 29892436<\/a><\/p>\n<p>Using synthetic oligonucleotides as standards in probe-based qPCR. <em>Biotechniques.<\/em> 2018 Apr; 64(4):177-179. <a href=\"https:\/\/pubmed.ncbi.nlm.nih.gov\/29661012\/\">PMID: 29661012<\/a><\/p>\n","protected":false},"excerpt":{"rendered":"<p>EB\u30a6\u30a4\u30eb\u30b9\uff08Epstein-Barr virus; EBV\uff09\u306f\u3001\u30d8\u30eb\u30da\u30b9\u30a6\u30a3\u30eb\u30b9\u79d1\u306b\u5c5e\u3059\u308b\u75c5\u539f\u6027\u30a6\u30a4\u30eb\u30b9\u306e\u4e00\u7a2e\u3067\u3042\u308a\u3001\u553e\u6db2\u3092\u4ecb\u3057\u3066\u611f\u67d3\u3092\u5e83\u3052\u3001\u4e3b\u306bB\u7d30\u80de\u306b\u611f\u67d3\u3057\u307e\u3059\u3002EBV\u306f95%\u4ee5\u4e0a\u306e\u5065\u5e38\u6210\u4eba\u304c\u611f\u67d3\u3057\u3001\u305d\u306e\u591a\u304f\u304c\u7d42\u751f\u5171\u751f\u7684\u306b\u4fdd\u6301\u3055\u308c\u308b\u8ab0\u3082\u304c\u611f\u67d3\u3057\u3066\u3044\u308b\u30a6\u30a4\u30eb\u30b9\u3067\u3042\u308b\u4e00\u65b9\u3001\u767a\u75c7\u306f\u7a00\u3067\u306f\u3042\u308a\u307e\u3059\u304c\u3001\u30d0\u30fc\u30ad\u30c3\u30c8\u30ea\u30f3\u30d1\u816b\u306a\u3069\u4e00\u90e8\u306e\u60aa\u6027\u30ea\u30f3\u30d1\u816b\u3084\u4e0a\u54bd\u982d\u304c\u3093\u306e\u767a\u751f\u3068\u95a2\u9023\u304c\u3042\u308b\u3053\u3068\u304c\u660e\u3089\u304b\u306b\u306a\u3063\u3066\u3044\u307e\u3059\u3002  [&hellip;]<\/p>\n","protected":false},"author":14,"featured_media":0,"parent":9901,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"","meta":{"_seopress_robots_primary_cat":"","_seopress_titles_title":"","_seopress_titles_desc":"","_seopress_robots_index":"","_mc_calendar":[],"footnotes":"","_wp_rev_ctl_limit":""},"class_list":["post-9905","page","type-page","status-publish","hentry"],"_links":{"self":[{"href":"http:\/\/cell.brc.riken.jp\/ja\/wp-json\/wp\/v2\/pages\/9905","targetHints":{"allow":["GET"]}}],"collection":[{"href":"http:\/\/cell.brc.riken.jp\/ja\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"http:\/\/cell.brc.riken.jp\/ja\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"http:\/\/cell.brc.riken.jp\/ja\/wp-json\/wp\/v2\/users\/14"}],"replies":[{"embeddable":true,"href":"http:\/\/cell.brc.riken.jp\/ja\/wp-json\/wp\/v2\/comments?post=9905"}],"version-history":[{"count":6,"href":"http:\/\/cell.brc.riken.jp\/ja\/wp-json\/wp\/v2\/pages\/9905\/revisions"}],"predecessor-version":[{"id":15603,"href":"http:\/\/cell.brc.riken.jp\/ja\/wp-json\/wp\/v2\/pages\/9905\/revisions\/15603"}],"up":[{"embeddable":true,"href":"http:\/\/cell.brc.riken.jp\/ja\/wp-json\/wp\/v2\/pages\/9901"}],"wp:attachment":[{"href":"http:\/\/cell.brc.riken.jp\/ja\/wp-json\/wp\/v2\/media?parent=9905"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}