{"id":34140,"date":"2018-08-08T01:00:00","date_gmt":"2018-08-09T04:34:09","guid":{"rendered":"http:\/\/www.05921.net\/index.php\/2018\/08\/09\/%e8%a1%8c%e4%b8%9a%e5%8a%a8%e6%80%81-%e5%8d%8e%e9%93%b6%e5%81%a5%e5%ba%b7%e5%85%8d%e7%96%ab%e7%bb%84%e5%ba%93%e6%b5%8b%e5%ba%8f%e6%8a%80%e6%9c%af%e9%87%8d%e7%a3%85%e5%8d%87%e7%ba%a7%ef%bc%81\/"},"modified":"2018-08-09T12:34:14","modified_gmt":"2018-08-09T04:34:14","slug":"%e8%a1%8c%e4%b8%9a%e5%8a%a8%e6%80%81-%e5%8d%8e%e9%93%b6%e5%81%a5%e5%ba%b7%e5%85%8d%e7%96%ab%e7%bb%84%e5%ba%93%e6%b5%8b%e5%ba%8f%e6%8a%80%e6%9c%af%e9%87%8d%e7%a3%85%e5%8d%87%e7%ba%a7%ef%bc%81","status":"publish","type":"post","link":"https:\/\/www.05921.net\/index.php\/34140\/","title":{"rendered":"\u884c\u4e1a\u52a8\u6001 | \u534e\u94f6\u5065\u5eb7\u514d\u75ab\u7ec4\u5e93\u6d4b\u5e8f\u6280\u672f\u91cd\u78c5\u5347\u7ea7\uff01\u53ef\u9ad8\u6548\u3001\u5feb\u901f\u5206\u6790\u5fae\u91cf\u6837\u672c"},"content":{"rendered":"
\n

\"\u884c\u4e1a\u52a8\u6001
<\/p>\n

\u4e3a\u62b5\u5fa1\u5404\u79cd\u5404\u6837\u7684\u75c5\u539f\u4f53\uff0c\u4eba\u7c7b\u6216\u54fa\u4e73\u52a8\u7269\u7684\u9002\u5e94\u6027\u514d\u75ab\u7cfb\u7edf\u62e5\u6709\u5927\u91cf\u7684T\u7ec6\u80de\u53d7\u4f53(TCRs)\u548cB\u7ec6\u80de\u53d7\u4f53(BCRs)\uff0c\u4e5f\u88ab\u79f0\u4e3a\u514d\u75ab\u7ec4\u5e93\u3002\u9ad8\u901a\u91cf\u6d4b\u5e8f\uff08NGS\uff09\u6280\u672f\u53ef\u5bf9T\u6216B\u7ec6\u80de\u514b\u9686\u8fdb\u884c\u6df1\u5165\u5206\u6790\uff0c\u6709\u6548\u8bc4\u4f30TCR\u6216BCR\u4e92\u8865\u51b3\u5b9a\u533a\u57df(CDR3)\u7684\u591a\u6837\u6027\uff0c\u5e76\u8bc4\u4f30\u514b\u9686\u7c7b\u578b\u7684\u6784\u6210\uff0c\u4f7fT\u6216B\u7ec6\u80de\u68c0\u6d4b\u8fbe\u5230\u524d\u6240\u672a\u6709\u7684\u7cbe\u7ec6\u5ea6\u3002\u514d\u75ab\u7ec4\u5e93NGS\u68c0\u6d4b\u4e3a\u8bc4\u4f30\u5065\u5eb7\u6216\u75be\u75c5\u72b6\u6001\u4e0b\u7684\u673a\u4f53\u9002\u5e94\u6027\u514d\u75ab\u7cfb\u7edf\u63d0\u4f9b\u4e86\u6280\u672f\u652f\u6301\u3002<\/strong><\/span>
<\/p>\n

\n
\n
\n
\n

\"\u884c\u4e1a\u52a8\u6001
<\/p>\n<\/section>\n<\/section>\n<\/section>\n

\n
\n
\n

\u76ee\u524d\uff0c\u73b0\u6709\u514d\u75ab\u7ec4\u5e93\u6280\u672f\u4e3b\u8981\u5206\u4e3a\u4e24\u7c7b\uff1a<\/span><\/p>\n

\n
\n
\n
\n
\n
\n
\n

 1. \u57fa\u4e8e\u591a\u91cdPCR\u7684\u6587\u5e93\u6784\u5efa\u65b9\u6848\uff1a\u4ee5DNA\u4e3a\u6a21\u677f\uff0c\u9488\u5bf9\u53ef\u53d8\u533a\uff08CDR3\uff09\u8bbe\u8ba1\u591a\u5bf9\u5f15\u7269\uff0c\u6269\u589e\u51faCDR3\u533a\u6bb5\uff0c\u518d\u8fdb\u884c\u6587\u5e93\u6784\u5efa\u5e76\u6d4b\u5e8f\uff1b<\/span><\/p>\n

2. \u57fa\u4e8e5’RACE\u7684\u6587\u5e93\u6784\u5efa\u65b9\u6848\uff1a\u4ee5mRNA\u4e3a\u6a21\u677f\uff0c\u5728\u6052\u5b9a\u533a\u8bbe\u8ba1\u5f15\u7269\u9006\u8f6c\u5f55\u6269\u51faTCR\u6216BCR\u7684\u5168\u957f\u5e8f\u5217\uff0c\u7ecf\u8fc7\u4e24\u8f6ePCR\u5bcc\u96c6\u8fd9\u4e00\u76ee\u7684\u5e8f\u5217\uff0c\u518d\u8fdb\u884c\u6587\u5e93\u6784\u5efa\u5e76\u6d4b\u5e8f\u3002<\/span><\/p>\n<\/section>\n<\/section>\n<\/section>\n<\/section>\n<\/section>\n<\/section>\n<\/section>\n

\u5c3d\u7ba1NGS\u6d4b\u5e8f\u6280\u672f\u53ef\u5feb\u901f\u4fbf\u6377\u5730\u5bf9\u514d\u75ab\u7ec4\u5e93\u8fdb\u884c\u5168\u666f\u5f0f\u626b\u63cf\uff0c\u4f46\u73b0\u6709\u65b9\u6cd5\u4ecd\u5b58\u5728\u660e\u663e\u7684\u4e0d\u8db3\u4e4b\u5904\u3002\u5982<\/span>\u57fa\u4e8e\u591a\u91cdPCR\u7684\u6587\u5e93\u6784\u5efa\u65b9\u6848\u540c\u65f6\u8fdb\u884c\u51e0\u5341\u79cd\u5f15\u7269\u7684\u6269\u589e\uff0c\u4f53\u7cfb\u5f02\u5e38\u590d\u6742\uff1b\u591a\u91cd\u5f15\u7269\u8bbe\u8ba1\u9700\u5df2\u77e5\u53c2\u8003\u5e8f\u5217\uff0c\u4e0d\u80fd\u5145\u5206\u6355\u6349\u5230\u4eba\u7c7b\u7b49\u4f4d\u57fa\u56e0\u7a81\u53d8\u5e8f\u5217\uff1b\u591a\u79cd\u5f15\u7269\u6269\u589e\u6548\u7387\u53ef\u80fd\u5b58\u5728\u504f\u501a\uff0c\u96be\u4ee5\u7b49\u6548\u6269\u589e\uff1b\u4e0e\u4f7f\u7528mRNA\u6784\u5efa\u6587\u5e93\u76f8\u6bd4\uff0cDNA\u6240\u9700\u7684PCR\u53cd\u5e94\u4f53\u7cfb\u8f83\u5927\uff0c\u6837\u672c\u8981\u6c42\u8f83\u9ad8\u3002\u8fd9\u4e9b\u4e0d\u8db3\u53ef\u80fd\u4f1a\u5bfc\u81f4\u5047\u9633\u6027\u589e\u52a0\u548c\u6269\u589e\u504f\u501a\uff0c\u4ece\u800c\u5bf9\u6570\u636e\u7ed3\u679c\u7684\u51c6\u786e\u6027\u9020\u6210\u6f5c\u5728\u5f71\u54cd\u3002<\/span><\/p>\n

\u4e3a\u6539\u8fdb\u4e0a\u8ff0\u514d\u75ab\u7ec4\u5e93\u6d4b\u5e8f\u65b9\u6cd5\uff0c\u534e\u94f6\u5065\u5eb7\u5bf9\u5df2\u6709\u76845’RACE\u6587\u5e93\u6784\u5efa\u6280\u672f\u8fdb\u884c\u4e86\u5347\u7ea7\uff0c\u63a8\u51fa\u4e86\u5168\u65b0\u7684\u514d\u75ab\u7ec4\u5e93\u6d4b\u5e8f\u6280\u672f\u670d\u52a1\u3002<\/span><\/strong><\/p>\n

\n
\n
\n
\n
\n
\n
\n

 <\/span>\u6280\u672f\u5347\u7ea7\u4e00\uff1a<\/strong><\/span><\/p>\n<\/section>\n<\/section>\n<\/section>\n<\/section>\n<\/section>\n<\/section>\n<\/section>\n

\n
\n
\n
\n
\n
\n
\n
\n

\u57fa\u4e8e5’RACE\u539f\u7406\uff0c\u5e76\u5728\u539f\u59cb\u6a21\u677f\u6269\u589e\u524d\u5f15\u5165UMI\uff08Unique Molecular Indices\uff0c\u5206\u5b50\u6761\u5f62\u7801\uff09\u8fdb\u884c\u6587\u5e93\u6784\u5efa\u3002\u5728\u6052\u5b9a\u533a\u5355\u5f15\u7269\u9006\u8f6c\u5f55\u6269\u589e\u51faCDR\u533a\u57df\u5168\u957f\u5e8f\u5217\u7684\u540c\u65f6\uff0c\u5f15\u5165UMI\u5e8f\u5217\uff0c\u53ef\u5bf9\u540e\u7eedPCR\u53cd\u5e94\u6216\u6d4b\u5e8f\u8fc7\u7a0b\u4e2d\u5f15\u5165\u7684\u9519\u8bef\u8fdb\u884c\u6709\u6548\u7ea0\u6b63\uff0c\u63d0\u9ad8\u51c6\u786e\u7387\u3002<\/strong><\/span><\/p>\n

\n
\n
\n
\n
\n
\n
\n

\u6280\u672f\u5347\u7ea7\u4e8c\uff1a<\/strong><\/span><\/p>\n<\/section>\n<\/section>\n<\/section>\n<\/section>\n<\/section>\n<\/section>\n<\/section>\n

\u5728\u9006\u8f6c\u5f55\u73af\u8282\uff0c\u8bbe\u8ba1\u9488\u5bf9\u5fae\u91cf\u6837\u54c1\uff08\u4f4e\u81f3100\u4e2a\u7ec6\u80de\uff09\u7684\u5b9e\u9a8c\u6d41\u7a0b<\/span><\/strong>\uff1a\u5373\u901a\u8fc7oligo dT\u78c1\u73e0\u6355\u83b7mRNA\uff0c\u4ee5\u78c1\u73e0\u4e0a\u7684oligo dT\u4e3a\u9006\u8f6c\u5f55\u5f15\u7269\u5408\u6210cDNA\uff0c\u5e76\u7eaf\u5316\u56de\u6536cDNA\uff0c\u8f83\u5e38\u89c4\u65b9\u6cd5\u53ef\u5927\u5927\u63d0\u9ad8\u6548\u7387\uff0c\u4fdd\u8bc1\u4e86\u4eba\u6216\u5c0f\u9f20\u6765\u6e90\u7684\u5fae\u91cf\u6837\u672c\u7684\u5b9e\u9a8c\u6210\u529f\u7387\u3002<\/span><\/p>\n

\n
\n
\n
\n

\u5168\u65b0\u514d\u75ab\u7ec4\u5e93\u6d4b\u5e8f\u6280\u672f\u670d\u52a1\u4eae\u70b9<\/span><\/strong><\/span><\/p>\n<\/section>\n<\/section>\n<\/section>\n<\/section>\n<\/section>\n<\/section>\n<\/section>\n<\/section>\n<\/section>\n<\/section>\n<\/section>\n<\/section>\n<\/section>\n<\/section>\n<\/section>\n

\n
\n
\n
\n
\n

1. \u53ef\u6ee1\u8db3\u5fae\u91cf\u6837\u672c\u5efa\u5e93\u7684\u5b9e\u9a8c\u8981\u6c42\uff08\u6700\u4f4e\u53ef\u81f3100\u4e2a\u7ec6\u80de\uff09\uff0c\u517c\u5bb9\u4eba\u548c\u5c0f\u9f20\u6837\u672c\u3002<\/span><\/strong><\/p>\n

\"\u884c\u4e1a\u52a8\u6001<\/p>\n

\u5fae\u91cfRNA\u8d77\u59cb\u5efa\u5e93\u6d41\u7a0b\u56fe<\/span><\/p>\n

2. \u4f4eRNA\u8d77\u59cb\u91cf\u7684\u6837\u672c\u4e5f\u80fd\u5feb\u901f\u8fbe\u5230\u6d4b\u5e8f\u9971\u548c\u671f\uff0c\u76f8\u540cRNA\u8d77\u59cb\u91cf\u4e0b\u7684\u6837\u672c\u5176\u76f8\u5173\u5ea6\u8f83\u9ad8\u3002<\/span><\/strong><\/p>\n

\"\u884c\u4e1a\u52a8\u6001<\/p>\n

\u540c\u4e00\u6837\u672cRNA\u8d77\u59cb\u91cf\u4e0d\u540c\u65f6\u5230\u8fbe\u6d4b\u5e8f\u9971\u548c\u671f\u6570\u636e\u7edf\u8ba1\u66f2\u7ebf\u56fe<\/span><\/p>\n

\"\u884c\u4e1a\u52a8\u6001<\/p>\n

\u540c\u4e00\u6837\u672cRNA\u8d77\u59cb\u91cf\u76f8\u540c\u65f6\u6837\u672c\u95f4\u7684\u76f8\u5173\u6027\u56fe<\/span><\/p>\n

\uff08A1\/A2\/A3\u4e3a\u8d77\u59cbRNA\u91cf\u4e3a1600ng; B1\/B2\/B3\u4e3a\u8d77\u59cb\u91cf\u4e3a400ng; C1\/C2\/C3\u4e3a\u8d77\u59cb\u91cf\u4e3a50ng\uff09<\/span><\/p>\n

3. \u57fa\u4e8e\u5f3a\u5927\u7684UMI\u6807\u8bb0\u4e0e\u6821\u6b63\u7b97\u6cd5\uff0c\u53ef\u6709\u6548\u7ea0\u6b63PCR\u4e0e\u6d4b\u5e8f\u8fc7\u7a0b\u5f15\u5165\u7684\u9519\u8bef\u4e0e\u504f\u501a\uff0c\u6781\u5927\u63d0\u9ad8\u51c6\u786e\u5ea6\u3002<\/span><\/strong><\/p>\n

\"\u884c\u4e1a\u52a8\u6001<\/p>\n

\u7ecf\u8fc7UMI\u7b97\u6cd5\u6821\u6b63\u540e\u7684\u90e8\u5206\u6d4b\u8bd5\u6570\u636e\u7edf\u8ba1\u8868<\/span><\/p>\n

4. \u5bf9\u6d4b\u5e8f\u4e0b\u673a\u6570\u636e\u8fdb\u884c\u591a\u7ef4\u5ea6\u5206\u6790\uff0c\u7ed9\u51fa\u6700\u5168\u9762\u7684\u6570\u636e\u7ed3\u679c\u62a5\u544a\uff0c\u5e76\u53ef\u5b9a\u5236\u4e2a\u6027\u5316\u5206\u6790\u670d\u52a1\u3002<\/span><\/strong><\/p>\n

\"\u884c\u4e1a\u52a8\u6001<\/p>\n

\u8fd1\u5e74\u6765\uff0c\u57fa\u4e8e\u514d\u75ab\u7ec4\u5e93\u591a\u6837\u6027\u53d8\u5316\u7279\u70b9\u7684\u751f\u7269\u6807\u5fd7\u7269\u53d1\u73b0\u3001\u80bf\u7624\u7b49\u75be\u75c5\u7597\u6548\u9884\u6d4b\u3001\u75be\u75c5\u7684\u6613\u611f\u6027\u548c\u62b5\u6297\u6027\u3001\u611f\u67d3\u6027\u75be\u75c5\u53ca\u75ab\u82d7\u7814\u7a76\u7b49\u65b9\u9762\u90fd\u53d6\u5f97\u4e86\u91cd\u8981\u8fdb\u5c55\u3002<\/span>\u514d\u75ab\u7ec4\u5e93\u66f4\u662f\u88ab\u79f0\u4e3a\u751f\u7269\u6807\u8bb0\u7269\u7684\u201c\u91d1\u77ff\u201d\u3002<\/span>\u968f\u7740\u514d\u75ab\u7ec4\u5e93\u68c0\u6d4b\u7814\u7a76\u7684\u4e0d\u65ad\u53d1\u5c55\u548c\u6210\u719f\uff0c\u8d8a\u6765\u8d8a\u591a\u7684\u6d4b\u5e8f\u6280\u672f<\/span>\u548c\u65b9\u6cd5\u88ab\u5de7\u5999\u6574\u5408\uff0c\u5404\u6d4b\u5e8f\u6280\u672f\u4e5f\u5728\u4e0d\u65ad\u5347\u7ea7\u4f18\u5316\uff0c\u5728\u89e3\u51b3\u7814\u7a76\u96be\u9898\u7684\u540c\u65f6\uff0c<\/span><\/span>\u62d3\u5bbd\u4e86\u4f20\u7edf\u80bf\u7624\u5b66\u3001\u514d\u75ab\u5b66\u7b49\u7684\u7814\u7a76\u89c6\u91ce\uff0c\u4e5f\u4e3a\u4e34\u5e8a\u75be\u75c5\u8bca\u65ad\u63d0\u4f9b\u4e86\u65b0\u53c2\u8003\u3002<\/span><\/p>\n<\/section>\n<\/section>\n<\/section>\n<\/section>\n<\/section>\n

\n
\n
\n

\u53c2\u8003\u6587\u732e\uff1a<\/span><\/p>\n

1.Abbas A K, et al. Cellular & Molecular Immunology 6e[J].<\/span><\/p>\n

2.Hou X L, et al. Current status and recent advances of next generation sequencing techniques in immunological repertoire[J]. Genes & Immunity, 2016, 17(3):153.<\/span><\/p>\n

3.Lefranc, et al. “IMGT\u00ae, the international ImMunoGeneTics information system\u00ae.” Nucleic acids research 37.suppl_1 (2008): D1006-D1012.<\/span><\/p>\n

4.Chunlin Wang, et al. High throughput sequencing reveals a complex pattern of dynamic interrelationships among human T cell subsets. PNAS, 2010, vol. 107:1518\u20131523.<\/span><\/p>\n

5.Jennifer Benichou, et al. Rep-Seq: uncovering the immunological repertoire through next-generation sequencing. Immunology, 2011, 135: 183\u2013191.<\/span><\/p>\n

6.Jeroen W J van Heijst, et al. Quantitative assessment of T cell repertoire recovery after hematopoietic stem cell transplantation. Nature Medicine, 2013,VOL.19: 372-379.<\/span><\/p>\n

7.Miran Jang, et al. Characterization of T cell repertoire of blood,tumor, and ascites in ovarian cancer patients using next generation sequencing. OncoImmunology, 2015, Vol.4:e1030561-1-10.<\/span><\/p>\n

8.Evaggelia Liaskou, et al. High-throughput T-cell receptor sequencing across chronic liver diseases reveals distinct disease-associated repertoires. Hepatology, 2015,1-10.<\/span><\/p>\n

9.Bates S T, et al. Global biogeography of highly diverse protistan communities in soil[J]. The ISME journal, 2013,7(3): 652-659.<\/span><\/p>\n

\"\u884c\u4e1a\u52a8\u6001<\/p>\n

\u4e86\u89e3\u66f4\u591a\u9ad8\u901a\u91cf\u6d4b\u5e8f\u76f8\u5173\u4fe1\u606f\uff0c\u8bf7\u54a8\u8be2\u534e\u94f6\u5065\u5eb7\u9ad8\u901a\u91cf\u68c0\u6d4b\u4e8b\u4e1a\u90e8\uff1a400-100-8786<\/span><\/p>\n<\/section>\n<\/section>\n<\/section>\n<\/section>\n

\u00b7 END \u00b7<\/span><\/strong><\/p>\n

\n
\n
\n
\n
\n
\n
\n
\n
\n
<\/polygon><\/polygon><\/polygon><\/polygon><\/polygon><\/polygon><\/g><\/rect><\/g><\/rect><\/g><\/g><\/svg><\/section>\n<\/section>\n<\/section>\n<\/section>\n<\/section>\n<\/section>\n
\n
\n
\n

\u6309\u8bc6\u522b\u4e0b\u65b9\u4e8c\u7ef4\u7801\u5173\u6ce8\u63a2\u57fa\uff0c\u70b9\u51fb\u201c<\/span><\/strong><\/span><\/strong>\u8bfe\u7a0b\u62a5\u540d<\/span><\/strong>\u201d\uff0c\u8fdb\u5165<\/span><\/strong><\/span><\/strong>\u201c<\/span><\/strong><\/span><\/strong><\/span><\/span>\u5fc5\u5b66\u6570\u636e\u5e93<\/span><\/strong>\u201d<\/span><\/strong><\/span><\/strong><\/span>\uff0c\u62a5\u540d\u89c2\u770b\u8bfe\u7a0b\uff01<\/span><\/strong><\/span><\/strong><\/span><\/p>\n

\n<\/section>\n<\/section>\n<\/section>\n

\n
\n
\n
\n
\n
<\/polygon><\/polygon><\/polygon><\/polygon><\/polygon><\/polygon><\/g><\/rect><\/g><\/rect><\/g><\/g><\/svg><\/section>\n<\/section>\n<\/section>\n<\/section>\n<\/section>\n<\/section>\n<\/section>\n<\/section>\n<\/section>\n<\/section>\n

\"\u884c\u4e1a\u52a8\u6001
<\/p>\n

\n

\u59cb\u53d1\u4e8e\u5fae\u4fe1\u516c\u4f17\u53f7\uff1a \u6d4b\u5e8f\u4e2d\u56fd <\/p>\n<\/blockquote>\n<\/div>\n","protected":false},"excerpt":{"rendered":"

\u4e3a\u62b5\u5fa1\u5404\u79cd\u5404\u6837\u7684\u75c5\u539f\u4f53\uff0c\u4eba\u7c7b\u6216\u54fa\u4e73\u52a8\u7269\u7684\u9002\u5e94\u6027\u514d\u75ab\u7cfb\u7edf\u62e5\u6709\u5927\u91cf\u7684T\u7ec6\u80de\u53d7\u4f53(TCRs)\u548cB\u7ec6\u80de\u53d7\u4f53(BCRs) […]<\/p>\n","protected":false},"author":1,"featured_media":34141,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":[],"categories":[2],"tags":[],"_links":{"self":[{"href":"https:\/\/www.05921.net\/index.php\/wp-json\/wp\/v2\/posts\/34140"}],"collection":[{"href":"https:\/\/www.05921.net\/index.php\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/www.05921.net\/index.php\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/www.05921.net\/index.php\/wp-json\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/www.05921.net\/index.php\/wp-json\/wp\/v2\/comments?post=34140"}],"version-history":[{"count":1,"href":"https:\/\/www.05921.net\/index.php\/wp-json\/wp\/v2\/posts\/34140\/revisions"}],"predecessor-version":[{"id":34155,"href":"https:\/\/www.05921.net\/index.php\/wp-json\/wp\/v2\/posts\/34140\/revisions\/34155"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/www.05921.net\/index.php\/wp-json\/wp\/v2\/media\/34141"}],"wp:attachment":[{"href":"https:\/\/www.05921.net\/index.php\/wp-json\/wp\/v2\/media?parent=34140"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.05921.net\/index.php\/wp-json\/wp\/v2\/categories?post=34140"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.05921.net\/index.php\/wp-json\/wp\/v2\/tags?post=34140"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}