{"id":3021,"date":"2024-06-25T23:48:14","date_gmt":"2024-06-25T15:48:14","guid":{"rendered":"https:\/\/opentrons.com.cn\/?post_type=knowledge2&#038;p=3021"},"modified":"2024-09-02T18:57:05","modified_gmt":"2024-09-02T10:57:05","slug":"syrnadvance-viralheot-2pt","status":"publish","type":"post","link":"https:\/\/opentrons.com.cn\/zh-TW\/news\/syrnadvance-viralheot-2pt\/","title":{"rendered":"\u4f7f\u7528RNAdvance Viral\u548cOT-2\u5e73\u53f0\uff0c\u9032\u884c\u9ad8\u6548\u4e14\u9ad8\u901a\u91cf\u7684SARS-CoV-2\u75c5\u6bd2RNA\u6aa2\u6e2c\u53ca\u5b9a\u5e8f\u7814\u7a76"},"content":{"rendered":"<!-- wp:paragraph -->\n<p>\u61c9\u7528\u4ecb\u7d392020\u5e74\u65b0\u51a0\u75c5\u6bd2\u8086\u8650\u5168\u7403\uff0c\u6fc0\u5316\u4e86\u4eba\u985e\u5c0d\u65bc\u81e8\u5e8a\u7814\u7a76\u76ee\u7684\u7684\u9ad8\u901a\u91cf RNA \u6aa2\u6e2c\u548c\u57fa\u56e0\u5b9a\u5e8f\u7684\u9700\u6c42\u3002\u50b3\u7d71\u624b\u52d5\u64cd\u4f5c\u65b9\u6848\u96e3\u4ee5\u63d0\u5347RNA \u6aa2\u6e2c\u7684\u6548\u7387\u3002\u5728\u672c\u9805\u7814\u7a76\u4e2d\uff0c\u6211\u5011\u5c55\u793a\u4e86\u5728 Opentrons OT-2 \u81ea\u52d5\u5316\u79fb\u6db2\u5de5\u4f5c\u7ad9\u4e0a\u4f7f\u7528 Beckman Coulter Life Sciences RNAd- vance Viral \u8a66\u5291\u76d2 (C63510) \u7684\u81ea\u52d5\u5316 SARS-CoV-2 RNA \u8403\u53d6\u5de5\u4f5c\u6d41\u7a0b\u3002 \u6211\u5011\u63d0\u4f9b\u7684\u5206\u6790\u6578\u64da\u986f\u793a\u4e86\u900f\u904e qRT-PCR \u5b9a\u91cf\u7684SARS-CoV-2 \u6aa2\u6e2c\u6975\u9650(LoD) \u548c\u5e8f\u5217\u6bd4\u5c0d\u6578\u64da\uff0c\u9019\u4e9b\u6578\u64da\u70ba\u81ea\u52d5\u5b9a\u5e8f\u6a23\u672c\u548c\u6587\u5eab\u88fd\u5099\u63d0\u4f9b\u4e86\u4f9d\u64da\uff0c\u4ee5\u4fbf\u5c0d\u7a81\u8b8a\u682a\u9032\u884c\u907a\u50b3\u9451\u5b9a\u3002<\/p>\n<!-- \/wp:paragraph -->\n\n<!-- wp:paragraph -->\n<p>RNAdvance \u75c5\u6bd2\u8a66\u5291\u76d2\u548c OT-2 \u5e73\u53f0\u6982\u8ff0RNAdvance Viral kit \u900f\u904e\u5229\u7528 SPRI \u9806\u78c1\u73e0\u6280\u8853\uff0c\u5f9e\u6a23\u672c\u4e2d\u9ad8\u6548\u63d0\u53d6\u75c5\u6bd2RNA\u3002 \u6211\u5011\u5728\u81ea\u52d5\u5316\u7684Opentrons \u5e73\u53f0\u4e0a\u4f7f\u7528\u4e86\u9019\u7a2e\u8a66\u5291\uff0c\u5f9e\u5408\u6210\u9f3b\u57fa\u8cea(110mM NaCl,1%w\/v\u7684\u8c6c\u80c3I \u578b\u9ecf\u6db2(Sigma M2378-100G) \u548c10\u03bcg\/mL w\/v\u4eba\u985e\u57fa\u56e0\u7d44DNA(Coriell NA12878), \u4ee590%v\/v \u7684TE\/SNM) \u6a23\u672c\u4e2d\u63d0\u53d6\u4e86\u4e0d\u540c\u6fc3\u5ea6\u7684\u71b1\u6ec5\u6d3b\u7684SARS-CoV-2(ATCC\u00b0VR-1986HKTM) \u3002 \u6bcf\u500b\u6fc3\u5ea6\u5305\u62ec20 \u500b\u6e2c\u8a66\u6a23\u672c\u30012\u500b\u967d\u6027\u5c0d\u7167\u548c2\u500b\u9670\u6027\u5c0d\u7167\u3002<\/p>\n<!-- \/wp:paragraph -->\n\n<!-- wp:paragraph -->\n<p>\u5de5\u4f5c\u6d41\u7a0b\u541e\u5410\u91cf\u57161\u986f\u793a\u4e86\u81ea\u52d5\u5316 RNAdvance \u75c5\u6bd2\u63d0\u53d6\u5de5\u4f5c\u6d41\u7a0b\u7684 OT-2\u7532\u677f\u4f48\u5c40\u3002\u6b64\u5de5\u4f5c\u6d41\u7a0b\u5f9e\u5411\u6a23\u54c1\u88c2\u89e3\u6db2\u4e2d\u6dfb\u52a0 Bind (BBB)\u958b\u59cb\uff0c\u6bcf\u500b\u6a23 \u54c1\u50c5\u9700\u8981\u6d88\u801710\u500b\u5438\u982d\uff0c\u53ef\u9748\u6d3b\u904b\u884c8\u500b\u6279\u6b21\u5927\u5c0f\u7684\u6700\u591a96\u500b\u6a23\u672c\u3002\u5831\u544a\u4e2d\u6240\u8ff0\u7684\u5de5\u4f5c\u6d41\u7a0b\u6301\u7e8c\u6642\u9593\u5305\u62ec\u5be6\u969b\u64cd\u4f5c\u6642\u9593\u3001\u6a23\u54c1\u88c2\u89e3\u6642\u9593\u548c\u512a\u5316\u5f8c\u6a23\u54c1\u7684\u904b\u884c\u6642\u9593(\u57162)\u3002\u4f7f\u7528RNAdvance Viral\u8a66\u5291\u76d2\u548cOT-2\u5e73\u53f0\uff0c\u8655\u740696\u500b\u6a23\u672c\u7684\u5de5\u4f5c\u6d41\u7a0b\u7e3d\u5171\u9700\u89813.5\u5c0f\u6642\uff0c\u60a8\u53ef\u4ee5\u8907\u88fd\u9023\u7d50\u5230\u700f\u89bd\u5668\uff0c\u67e5\u770b\u7279\u5b9a\u5354\u8b70\u6d41\u7a0b\uff1a https:\/\/protocols.opentrons.com\/protocol\/bc- rnadvance-viral\u3002\u60a8\u53ef\u4ee5\u6839\u64da\u60a8\u7684\u9700\u6c42\u9748\u6d3b\u5730\u6539\u8b8a\u65b9\u6cd5\u8b8a\u91cf\uff0c\u4ee5\u7372\u5f97\u60a8\u6240\u9700\u7684\u6279\u6b21\u5927\u5c0f\u3001\u6a23\u54c1\u8f38\u5165\u548c\u6d17\u812b\u9ad4\u7a4d\u7684\u6240\u9700\u7d50\u679c\u3002<\/p>\n<!-- \/wp:paragraph -->\n\n<!-- wp:image {\"id\":3022,\"sizeSlug\":\"full\",\"linkDestination\":\"none\"} -->\n<figure class=\"wp-block-image size-full\"><img src=\"https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-81.png\" alt=\"OT-2  \u5b9e\u9a8c\u7532\u677f\u5e03\u5c40\" class=\"wp-image-3022\"><figcaption class=\"wp-element-caption\"><strong>\u57161:\u914d\u5099\u6a21\u7d44\u3001\u5be6\u9a57\u8017\u6750\u53ca<\/strong>&nbsp;&nbsp;<strong>RNAdvance &nbsp;&nbsp;&nbsp;Viral &nbsp;\u8a66\u5291\u7684<\/strong>&nbsp;&nbsp;<strong>Opentrons &nbsp;&nbsp;&nbsp;OT-2 &nbsp;\u5be6\u9a57\u7532\u677f\u4f48\u5c40<\/strong><\/figcaption><\/figure>\n<!-- \/wp:image -->\n\n<!-- wp:paragraph -->\n<p>\u6b64\u5de5\u4f5c\u6d41\u7a0b\u5728 &nbsp;OT-2&nbsp;\u4e0a\u662f\u53ef\u4ee5\u5168\u81ea\u52d5\u904b\u4f5c&nbsp;&nbsp;\u7684\uff0c\u6bcf\u500b\u6a23\u672c\u4f7f\u752810\u500b\u5438\u982d\uff0c\u4e26\u5f9e\u52a0\u5165Bind(BBB) &nbsp;&nbsp;&nbsp;\u958b\u59cb\u3002\u5be6\u9a57\u7532\u677f\u4e0a\u8017\u6750\u5167\u5bb9\u6709\uff1a\u6700\u591a10\u500b\u5438\u982d\u67b6\u30011\u500b&nbsp;NEST&nbsp;\u55ae\u5b54\u5132\u6db2\u69fd\u30012\u500b&nbsp;NEST 12&nbsp;\u5b54\u5132\u6db2\u69fd\u30011\u500bNEST&nbsp;0.1mL&nbsp;PCR\u677f\u53ca1\u500b&nbsp;NEST &nbsp;2mL&nbsp;\u6df1\u5b54\u677f\uff0c\u6a21\u7d44\u5305\u62ec\u4e00\u500b\u78c1\u73e0\u7d14\u5316\u6a21\u7d44\u548c\u4e00\u500b\u6eab\u63a7\u6a21\u7d44\uff0c\u914d\u5099\u4e00\u500b&nbsp;P300&nbsp;\u591a\u901a\u9053\u79fb\u6db2\u5668\u3002\u6240\u6709\u7684&nbsp;NEST \u8017\u6750\u548c\u6a21\u7d44\u90fd\u7531&nbsp;Opentrons \u63d0\u4f9b\u3002<\/p>\n<!-- \/wp:paragraph -->\n\n<!-- wp:image {\"id\":3023,\"sizeSlug\":\"full\",\"linkDestination\":\"none\"} -->\n<figure class=\"wp-block-image size-full\"><img src=\"https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-82.png\" alt=\"\u5b9e\u9a8c\u6570\u636e\" class=\"wp-image-3023\"><\/figure>\n<!-- \/wp:image -->\n\n<!-- wp:image {\"id\":3024,\"sizeSlug\":\"full\",\"linkDestination\":\"none\"} -->\n<figure class=\"wp-block-image size-full\"><img src=\"https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-83.png\" alt=\"\u5b9e\u9a8c\u6570\u636e\" class=\"wp-image-3024\"><\/figure>\n<!-- \/wp:image -->\n\n<!-- wp:paragraph -->\n<p>\u57162:\u5728OT-2\u5e73\u53f0\u4e0a\u4f7f\u7528RNAdvance Viral\u8a66\u5291\u76d2\u7684\u541e\u5410\u91cf\u72c0\u6cc1\u6574\u500b\u5de5\u4f5c\u6d41\u7a0b\u904b\u4f5c\u6240\u9700\u7684\u6642\u9593\u5305\u542b3\u500b\u90e8\u5206\uff1a\u64cd\u4f5c\u6642\u9593\u3001\u88c2\u89e3\u6642\u9593\u548c\u904b\u884c\u6642\u9593\u3002\u64cd\u4f5c\u6642\u9593\u5305\u62ec\u6a23\u54c1\u548cOT-2\u7684\u6e96\u5099\u5de5\u4f5c\u3002\u904b\u884c\u6642\u9593\u6307\u7684\u662f OT-2\u5f9e\u958b\u59cb\u5230\u5b8c\u6210\u6574\u500b\u5354\u8b70\u6240\u9700\u7684\u6642\u9593\u3002 A)\u8655\u74068\u500b\u6a23\u672c\u9700\u898145\u5206\u9418\uff0c\u8655\u740696\u500b\u6a23\u672c\u9700\u89812\u5c0f\u664238\u5206\u3002 B) \u5716\u5f62\u5316\u986f\u793a\u4e86\u4e0d\u540c\u6a23\u54c1\u6578\u91cf\u4e0b\u7684\u904b\u8f49\u8655\u7406\u6642\u9593\u3002<\/p>\n<!-- \/wp:paragraph -->\n\n<!-- wp:paragraph -->\n<p>RNAdvance \u75c5\u6bd2\u8a66\u5291\u76d2\u6aa2\u6e2c\u9650\u5236\u6aa2\u6e2c\u9650\u5236(LoD)\u662f\u7528\u65bc\u78ba\u5b9aOT-2\u5e73\u53f0\u4e0a\u81ea\u52d5\u5316RNAdvance Viral\u8403\u53d6\u6548\u7387\u7684\u6307\u6a19\uff0c\u5b83\u8868\u793a\u80fd\u5920\u5728\u226595%\u7684\u91cd\u8907\u6e2c\u91cf\u4e2d\u6aa2\u6e2c\u5230\u7684\u75c5\u6bd2\u62f7\u8c9d\u6578 \u7684\u6700\u4f4e\u6fc3\u5ea6\u3002\u6211\u5011\u4f7f\u75282019-nCoV CDC N1\u548cRP\u5f15\u5b50\u548c\u63a2\u91dd(2019-nCoV CDC EUA kit,Integrated DNA Technologies (IDT)) \u9032\u884c\u5075\u6e2c\uff0cPCR \u6df7\u5408\u6db2\u548c\u9175\u7d20\u4f7f\u7528Luna\u00b0\u901a\u7528\u63a2\u91dd\u4e00\u6b65RT-q England BioLabs),PCR\u53cd\u61c9\u5728PCRmax ECO48\u5373\u6642PCR \u7cfb\u7d71\u4e0a\u9032\u884c\u3002\u5728SARS-CoV-2 \u6a23\u672c (n=20) \u548c\u967d\u6027\u5c0d\u7167\u6a23\u672c (n=2) \u4e2d\u6aa2\u6e2c\u5230\u75c5\u6bd2RNA (\u62f7\u8c9d\u6578\/\u03bcL), \u4f46\u5728\u9670\u6027\u5c0d\u7167\u6a23\u672c (n=2) \u4e2d\u672a\u6aa2\u6e2c\u5230\u3002\u6839\u64da\u88681\u7684\u7e3d\u7d50\uff0c\u5206\u6790\u7bc4\u570d\u5167\u7684\u6240\u6709\u5c0d\u6bd4\u967d\u6027\u6a23\u672c\u5747\u88ab\u78ba\u5b9a\u70ba\u967d\u6027\uff0c\u5176\u4e2d\u6700\u4f4e\u6fc3\u5ea6\u7684\u75c5\u6bd2RNA \u62f7\u8c9d\u6578\u70ba1\u62f7\u8c9d\/\u03bcL\u3002 PCR \u53c3\u6578\u5982\u4e0b\u3002<\/p>\n<!-- \/wp:paragraph -->\n\n<!-- wp:image {\"id\":3025,\"sizeSlug\":\"full\",\"linkDestination\":\"none\"} -->\n<figure class=\"wp-block-image size-full\"><img src=\"https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-84.png\" alt=\"\u5b9e\u9a8c\u6570\u636e\" class=\"wp-image-3025\"><\/figure>\n<!-- \/wp:image -->\n\n<!-- wp:paragraph -->\n<p>\u88681:\u4f7f\u7528OT-2 \u5e73\u53f0\u548c RNAdvance Viral \u8a66\u5291\u76d2\u6aa2\u6e2c SARS-CoV-2 \u76ee\u6a19 N1 \u7684\u75c5\u6bd2\u9748\u654f\u5ea6\u3002 1 copy\/\u03bcL \u662f\u80fd\u5920\u5075\u6e2c\u5230\u226595%\u6a23\u672c\u7684\u6700\u4f4e\u6fc3\u5ea6\uff0c\u88ab\u5b9a\u7fa9\u70ba\u5075\u6e2c\u9650\u5236 (Limit of Detection,LoD)<\/p>\n<!-- \/wp:paragraph -->\n\n<!-- wp:paragraph -->\n<p>\u70ba\u4e86\u7814\u7a76 RNAdvance Viral\u5728 OT-2 \u4e0a\u7684\u81ea\u52d5\u5316\u6027\u80fd\uff0c\u6211\u5011\u5c07850 copy\/\u03bcL \u6fc3\u5ea6\u7684\u71b1\u6ec5\u6d3b SARS-CoV-2 \u75c5\u6bd2\u52a0\u5165\u5408\u6210\u9f3b\u8154\u57fa\u8cea\u4e2d\uff0c\u4e26\u8403\u53d6RNA\u3002 \u91dd\u5c0dN1\u548cRP, \u78ba\u5b9a\u4e86\u5747\u503cCt\u503c\u3001\u6a19\u6e96\u5dee\u3001\u8b8a\u7570\u4fc2\u6578 (CV%) \u548c\u6aa2\u6e2c\u7387\u7b49\u898f\u683c\u53c3\u6578(\u57163A)\u3002\u5728\u5be6\u9a57\u6a23\u672c(n=20)\u3001 \u967d\u6027\u5c0d\u7167\u6a23\u672c(n=2) \u548c\u9670\u6027\u5c0d\u7167\u6a23\u672c(n=2) \u4e2d\uff0c\u5c0dN1 \u7684\u64f4\u589e\u66f2\u7dda\u9032\u884c\u5b9a\u91cf\u5206\u6790(\u57163B), \u540c\u6642\u5c0d\u5305\u62ec\u5c0d\u7167\u6a23\u672c\u5728\u5167\u7684\u6a23\u54c1\u4e2d\u7684RP\u9032\u884c\u5b9a\u91cf\u5206\u6790(\u57163C)\u3002<\/p>\n<!-- \/wp:paragraph -->\n\n<!-- wp:image {\"id\":3026,\"sizeSlug\":\"full\",\"linkDestination\":\"none\"} -->\n<figure class=\"wp-block-image size-full\"><img src=\"https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-85.png\" alt=\"\u5b9e\u9a8c\u7ed3\u679c\u8d8b\u52bf\u56fe\" class=\"wp-image-3026\"><\/figure>\n<!-- \/wp:image -->\n\n<!-- wp:paragraph -->\n<p>DNA \u5b9a\u5e8f\u8207\u6578\u64da\u5206\u6790\u70ba\u4e86\u9032\u4e00\u6b65\u4e86\u89e3\u5b9a\u5e8f\u6027\u80fd\uff0c\u6211\u5011\u4f7f\u7528Swift 2S Turbo DNA\u6587\u5eab\u88fd\u5099\u8a66\u5291\u76d2\u548c Swift Unique Dual Indexing \u5f15\u5b50\u677f\u9032\u884c N1\u64f4\u589e\u3002\u95dc\u65bc850 copy\/\u03bcl \u6a23\u672c\u7684\u8a73\u7d30\u8cc7\u6599\u548c\u5e8f\u5217\u6bd4\u5c0d\u7d50\u679c\u53ef\u4ee5\u5728\u57164 \u4e2d\u627e\u5230\u3002\u6211\u5011\u89c0\u5bdf\u5230\u5e73\u5747\u8986\u84cb\u7387\u70ba 455x, \u4e14\u5e73\u5747\u8b80\u6578\u670991%\u8207\u53c3\u8003\u57fa\u56e0\u7d44 SARS-CoV-2 \u5206\u96e2\u81eaWuhan-Hu-1(NC_045512.2) \u5c0d\u9f4a\u3002\u8b80\u53d6\u9577\u5ea6\u8207 N1\u64f4\u589e\u5340\u57df\u76f8\u5339\u914d\uff0c\u5927\u7d04\u70ba72\u500b\u9e7c\u57fa\u3002\u6839\u64daSwift 2s Turbo DNA \u6587\u5eab\u88fd\u5099\u8a66\u5291\u76d2\u88fd\u9020\u5546\u7684\u8aaa\u660e\uff0c\u9810\u671f\u63d2\u5165\u7247\u6bb5\u5927\u5c0f\u7d04\u70ba 200bp\u3002 \u5b9a\u5e8f\u662f\u4f7f\u7528 lllumina MiSeq \u9032\u884c\u7684\uff0c\u4e26\u5c07\u7be9\u9078\u8cc7\u6599\u4e0a\u50b3\u81f3llluminaBaseSpace\u3002\u5728 Galaxy \u4e2d\u4f7f\u7528 Bowtie2 \u78ba\u5b9a\u4e86\u8986\u84cb\u5ea6\uff0c\u4e26\u4f7f\u7528 Integrative Genome Viewer(IGV) \u9032\u884c\u8996\u89ba\u5316\u3002 NC_045512.2,SAR2-COV-2, \u5206\u96e2\u6e90\u81eaWuhan-Hu-1 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