{"id":88,"date":"2010-06-25T08:54:09","date_gmt":"2010-06-25T00:54:09","guid":{"rendered":"http:\/\/pckimo.com\/wordpress\/archives\/88"},"modified":"2010-06-25T09:04:59","modified_gmt":"2010-06-25T01:04:59","slug":"%e7%b4%b0%e8%aa%aa%e8%8f%9c%e8%8a%b1%e7%9a%84%e6%aa%a2%e9%a9%97%e8%88%87%e5%88%86%e6%9e%90","status":"publish","type":"post","link":"http:\/\/pckimo.com\/wordpress\/archives\/88","title":{"rendered":"\u7d30\u8aaa\u83dc\u82b1\u7684\u6aa2\u9a57\u8207\u5206\u6790"},"content":{"rendered":"<p>\u4e00\u822c\u5178\u578b\u7684\u83dc\u82b1\u53ef\u8996\u75c5\u7076\u7684\u72c0\u6cc1\uff0c\u7531\u76ee\u8996\u8fa8\u8a8d\uff0c\u4f46\u6709\u7591\u616e\u6642\uff0c\u4ecd\u9700\u9760\u6aa2\u9a57\u5206\u6790<\/p>\n<p>\u6709\u4ee5\u4e0b\u591a\u7a2e\u7684\u83dc\u82b1\u78ba\u8a3a\u65b9\u6cd5\uff0c\u4ee5\u75c5\u7406\u5207\u7247\u5f8c\u7684\u6aa2\u67e5\u624d\u80fd\u767e\u5206\u4e4b\u767e\u78ba\u5b9a\u70baHPV \u611f\u67d3\uff0c\u5c24\u4ee5\u7528PCR\u65b9\u6cd5\u6aa2\u6e2cHPV\u6bd4\u6838\u9178\u96dc\u4ea4\u65b9\u6cd5\u512a\u8d8a\u3002\u5176\u654f\u611f\u6027\u9ad8\uff0c\u6aa2\u6e2cHPV\u5be6\u9a57\u5468\u671f\u77ed\u3001\u7c21\u4fbf\u5feb\u901f\u3002<br \/>\n\u4ee5\u4e0b\uff1a<br \/>\n\u4e00\u3001\u918b\u9178\u767d\u8a66\u9a57<span style=\"background-color: #ccffff\">\uff08\u5047\u967d\u6027\u8f03\u5e38\u898b\uff09<\/span><\/p>\n<p>\u3000\u3000\u75283-5%\u918b\u9178\u5916\u5857\u75a3\u9ad42-5\u5206\u9418\uff0c\u75c5\u7ac8\u90e8\u4f4d\u8b8a\u767d\u7a0d\u9686\u8d77\uff0c\u809b\u9580\u75c5\u640d\u53ef\u80fd\u9700\u898115\u5206\u9418\u3002\u672c\u8a66\u9a57\u7684\u539f\u7406\u662f\u86cb\u767d\u8cea\u8207\u9178\u51dd\u56fa\u8b8a\u767d\u7684\u7d50\u679c\uff0cHPV\u611f\u67d3\u7d30\u80de\u7522\u751f\u7684\u89d2\u86cb\u767d\u8207\u6b63\u5e38\u7684\u672a\u611f\u67d3\u4e0a\u76ae\u7d30\u80de\u7522\u751f\u7684\u4e0d\u540c\uff0c\u53ea\u6709\u524d\u8005\u624d\u80fd\u88ab\u918b\u9178\u812b\u8272\u3002\u918b\u9178\u767d\u8a66\u9a57\u6aa2\u6e2cHPV\u7684\u654f\u611f\u6027\u5f88\u9ad8\uff0c\u5b83\u6bd4\u5e38\u898f\u6aa2\u6e2c\u89c0\u5bdf\u7d44\u7e54\u5b78\u8b8a\u5316\u9084\u597d\u3002\u4f46\u5076\u723e\u5728\u4e0a\u76ae\u589e\u539a\u6216\u5916\u50b7\u64e6\u7834\u75c5\u4f8b\u4e2d\u51fa\u73fe\u5047\u967d\u6027\u3001\u5047\u967d\u6027\u8b8a\u767d\u8de1\u8c61\u986f\u5f97\u754c\u9650\u4e0d\u6e05\u548c\u4e0d\u898f\u5247\u3002\u7f8e\u570bCDC\u63d0\u793a\uff0c\u918b\u9178\u767d\u8a66\u9a57\u4e26\u4e0d\u662f\u7279\u7570\u8a66\u9a57\uff0c\u4e14\u5047\u967d\u6027\u8f03\u5e38\u898b\u3002<\/p>\n<p>\u3000\u3000\u4e8c\u3001\u514d\u75ab\u7d44\u7e54\u5b78\u6aa2\u67e5<!--more--><\/p>\n<p>\u3000\u3000\u5e38\u7528\u904e\u6c27\u5316\u7269\u9176\u6297\u904e\u6c27\u5316\u7269\u9176\u65b9\u6cd5\uff08\u5373PAP\uff09\uff0c\u986f\u793a\u6fd5\u75a3\u5167\u7684\u75c5\u6bd2\u86cb\u767d\uff0c\u4ee5\u8b49\u660e\u75a3\u640d\u5bb3\u4e2d\u6709\u75c5\u6bd2\u6297\u539f\u3002HPV\u86cb\u767d\u967d\u6027\u6642\uff0c\u5c16\u92b3\u6fd5\u75a3\u7684\u6dfa\u8868\u4e0a\u76ae\u7d30\u80de\u5167\u53ef\u51fa\u73fe\u6de1\u7d05\u8272\u7684\u5f31\u967d\u6027\u53cd\u61c9\u3002<br \/>\n\u3000<br \/>\n\u3000\u3000\u4e09\u3001\u7d44\u7e54\u5316\u5b78\u6aa2\u67e5<\/p>\n<p>\u3000\u3000\u53d6\u5c11\u91cf\u75c5\u640d\u7d44\u7e54\u5236\u6210\u5857\u7247\uff0c\u7528\u7279\u7570\u6297\u4eba\u985e\u4e73\u982d\u7624\u75c5\u6bd2\u7684\u6297\u9ad4\u4f5c\u67d3\u8272\u3002\u5982\u75c5\u640d\u4e2d\u6709\u75c5\u6bd2\u6297\u539f\uff0c\u5247\u6297\u539f\u6297\u9ad4\u7d50\u5408\u3002\u5728\u904e\u6c27\u5316\u7269\u9176\u6297\u904e\u6c27\u5316\u7269\u9176\uff08PAP\uff09\u65b9\u6cd5\u4e2d\uff0c\u6838\u53ef\u88ab\u67d3\u6210\u7d05\u8272\u3002\u6b64\u6cd5\u7279\u7570\u6027\u5f37\u4e14\u8f03\u8fc5\u901f\uff0c\u5c0d\u8a3a\u65b7\u6709\u5e6b\u52a9\u3002<br \/>\n\u3000<br \/>\n\u3000\u3000\u56db\u3001\u75c5\u7406\u6aa2\u67e5<\/p>\n<p>\u3000\u3000\u4e3b\u8981\u70ba\u89d2\u5316\u4e0d\u5168\uff0c\u68d8\u5c64\u9ad8\u5ea6\u80a5\u539a\uff0c\u4e73\u982d\u7624\u6a23\u589e\u751f\uff0c\u8868\u76ae\u7a81\u589e\u539a\uff0c\u5ef6\u9577\uff0c\u5176\u589e\u751f\u7a0b\u5ea6\u53ef\u4f3c\u5047\u6027\u4e0a\u76ae\u7624\u6a23\u3002\u523a\u7d30\u80de\u548c\u57fa\u5e95\u7d30\u80de\u4e26\u6709\u76f8\u7576\u6578\u91cf\u7684\u6838\u5206\u88c2\u3001\u9817\u4f3c\u764c\u8b8a\u3002\u4f46\u7d30\u80de\u6392\u5217\u898f\u5247\uff0c\u4e14\u589e\u751f\u4e0a\u76ae\u548c\u771f\u76ae\u4e4b\u9593\u754c\u9650\u6e05\u695a\u3002\u5176\u7279\u9ede\u70ba\u7c92\u5c64\u548c\u523a\u5c64\u4e0a\u90e8\u7d30\u80de\u6709\u660e\u986f\u7684\u7a7a\u6ce1\u5f62\u6210\u3002\u6b64\u7a2e\u7a7a\u6ce1\u7d30\u80de\u8f03\u6b63\u5e38\u5927\uff0c\u80de\u6f3f\u8457\u8272\u6de1\u3001\u4e2d\u592e\u6709\u5927\u800c\u5713\uff0c\u6df1\u55dc\u583f\u6027\u7684\u6838\u3002\u901a\u5e38\u771f\u76ae\u6c34\u816b\u3001\u6bdb\u7d30\u8840\u7ba1\u64f4\u5f35\u4ee5\u53ca\u5468\u570d\u8f03\u81f4\u5bc6\u7684\u6162\u6027\u708e\u6027\u6d78\u6f64\u3002Bushke-loewenstein\u5de8\u5927\u578b\u5c16\u92b3\u6fd5\u75a3\uff0c\u8868\u76ae\u6975\u5ea6\u5411\u4e0b\u751f\u9577\uff0c\u4ee3\u66ff\u4e86\u5176\u4e0b\u9762\u7684\u7d44\u7e54\u3001\u6613\u8207\u9c57\u72c0\u7d30\u80de\u76f8\u6df7\uff0c\u6545\u9808\u591a\u6b21\u6d3b\u6aa2\u3002\u82e5\u6709\u7de9\u6162\u767c\u5c55\u4e4b\u50be\u5411\uff0c \u5247\u70ba\u4e00\u7a2e\u4f4e\u5ea6\u60e1\u8b8a\u7684\u904e\u7a0b\uff0c\u5373\u6240\u8b02\u75a3\u72c0\u764c\u3002<\/p>\n<p>\u3000\u3000\u4e94\u3001\u57fa\u56e0\u8a3a\u65b7<\/p>\n<p>\u3000\u3000\u8fc4\u4eca\uff0cHPV\u96e3\u65bc\u7528\u50b3\u7d71\u7684\u75c5\u6bd2\u57f9\u990a\u53ca\u8840\u6e05\u5b78\u6280\u8853\u6aa2\u6e2c\uff0c\u4e3b\u8981\u5be6\u9a57\u8a3a\u65b7\u6280\u8853\u662f\u6838\u9178\u96dc\u4ea4\u3002\u8fd1\u5e74\u4f86\u767c\u5c55\u7684PCR\u65b9\u6cd5\u5177\u6709\u7279\u7570\u3001\u654f\u611f\u3001\u7c21\u4fbf\u3001\u5feb\u901f\u7b49\u512a\u9ede\uff0c\u70baHPV\u6aa2\u6e2c\u958b\u8f9f\u4e86\u65b0\u9014\u5f91\u3002 \u3000\u3000\uff08\u4e00\uff09\u6a19\u672c\u7684\u91c7\u96c6\u53ca\u8655\u7406<\/p>\n<p>\u3000\u30001\uff0e\u6a19\u672c\u7684\u91c7\u96c6\u548c\u9810\u8655\u7406\uff1a\u7528\u522e\u677f\u6216\u751f\u7406\u9e7d\u6c34\u6d78\u6f64\u7684\u68c9\u68d2\u5f9e\u9670\u9053\u548c\u5bae\u9838\u5916\u53e3\u53d6\u5206\u6ccc\u7269\u548c\u7d30\u80de\u3002\u5728\u4f5c\u7d30\u80de\u5b78\u6aa2\u67e5\u7684\u540c\u6642\uff0c\u5c07\u6a19\u672c\u653e\u51655ml\u542b\u67090.05%\u786b\u67f3\u6c5e\u7684PBS\u4e2d\uff0c\u7528PBS\u96e2\u5fc3\uff083000g\uff0c10min\uff09\u6d17\u6ecc2\u6b21\uff0c\u6c88\u7a4d\u7d30\u80de\u91cd\u61f8\u65bc1mlPBS\u4e2d\uff0c\u53d60.5ml\u7d30\u80de\u61f8\u6db2\u62bd\u63d0DNA\u3002<\/p>\n<p>\u3000\u30002\uff0e\u6a19\u672c\u6838\u9178\u7684\u63d0\u53d6\uff1a\u63091\u9ad4\u7a4d\u7d30\u80de\u61f8\u6db2\u52a010\u500d\u9ad4\u7a4d\u7684\u7d30\u80de\u88c2\u89e3\u6db2\uff0810mmol\/L Tris-HCl\uff0cpH 7.4\uff0c10mmol\/L EDTA\uff0c150mmol\/L NaCl\uff0c0.4%SDS\uff0c1.0mg\/ml\u86cb\u767d\u9176K\uff0937\u2103\u4e0b\u8655\u7406\u904e\u591c;\u4e14\u7b49\u9ad4\u7a4d\u915a\/\u6c2f\u4eff(1:1)\uff0c\u6c2f\u4eff\/\u7570\u620a\u9187\uff0824:1\uff09\u5404\u62bd\u63d02\u6b21\uff1b\u52a01\/10\u9ad4\u7a4d3mol\/L NaAc\uff08pH 5.2\uff09\u53ca2.5\u500d\u9ad4\u7a4d\u7121\u6c34\u4e59\u9187\u7f6e-20\u2103 2h\u6216\u904e\u591c\u6c88\u6fb1DNA\uff1b\u52a01\u9ad4\u7a4d\u4e59\u9187\u6d17\u6ecc1\u6b21\uff1b\u752860&mu;l\u542bRNA\u9176(100&mu;g\/ml)\u7684TE\u6db2(10mmol\/L Tris-HCl\uff0cpH 8.0\uff0c1.0mmol\/L EDTA)\u6eb6\u89e3DNA\uff0c37\u2103\u6eab\u80b230min\u3002<\/p>\n<p>\u3000\u3000\uff08\u4e8c\uff09PCR\u64f4\u589e<\/p>\n<p>\u3000\u30001\uff0e \u5f15\u7269\u8a2d\u8a08\u548c\u5408\u6210\uff1aHPV\u57fa\u56e0\u7d44\u53ef\u5206\u70ba\u65e9\u671f\u5340\uff08E\uff09\u548c\u665a\u671f\u5340\uff08L\uff09\uff0c\u6bcf\u5340\u542b\u4e00\u7cfb\u5217\u958b\u653e\u8b80\u78bc\u6846\u67b6\uff08ORF\uff09\u3002\u5e8f\u5217\u5206\u6790\u8868\u660e\uff0c\u5404\u578bHPV\u7684\u975e\u7de8\u78bc\u5340\u53caE1\u3001E6\u3001E7\u548cL1\u5340\u5747\u6709\u4fdd\u5b88\u5e8f\u5217\u3002Manos\u7b49\u5f9eHPVL1\u5340\u4e2d\u9078\u64c7\u4fdd\u5b88\u5e8f\u5217\u8a2d\u8a08\u5408\u6210\u5f15\u7269MY11\u548cMY09\u898b\u88681\uff0c\u8a72\u5f15\u7269\u8207HPV 6\u300111\u300116\u300118\u53ca33\u578b\u6709\u4e92\u88dc\u5e8f\u5217\uff0c\u4e5f\u53ef\u64f4\u589e\u5176\u5b83\u578bHPV\u3002<\/p>\n<p>\u3000\u3000Manos\u7b49\u8a2d\u8a08\u7684HPVL1\u901a\u7528\u5f15\u7269<\/p>\n<p>\u3000\u3000MY11\uff1aGCMCAGGGWCATAAYAATGG<\/p>\n<p>\u3000\u3000MY09\uff1aCGTCCMARRGGAWACTGATC<\/p>\n<p>\u3000\u30002\uff0ePCR\u53cd\u61c9\u8a66\u5291\uff1aTaq DNA\u805a\u5408\u9176\uff082U\/ml\uff09\u300110mmol\/L dNTP\u8caf\u5099\u6db2\uff08dATP\u3001dCTP\u3001dGTP\u3001dTTP\u540410mmol\/L\uff09\uff0c10&times;PCR\u7de9\u6c96\u6db2(500mmol KCl\u300140mmol\/L MgCl2\u3001100mmol\/L Tris-HCl\u3001pH 8.5)\uff0c100&mu;mol\/L MY11\u548cMY09\u8caf\u5099\u6db2\uff0c\u6ec5\u83cc\u7684\u73bb\u7483\u84b8\u993e\u5668\u5236\u5099\u7684\u84b8\u993e\u6c34\u3002<\/p>\n<p>\u3000\u30003\uff0ePCR\u64f4\u589e\u65b9\u6cd5\u548c\u7a0b\u5e8f\uff1a\u4ee5100&mu;l PCR\u53cd\u61c9\u6db2,\u7528\u7121\u83cc0.5ml\u77fd\u5316\u5851\u6599\u96e2\u5fc3\u7ba1\u70ba\u53cd\u61c9\u7ba1\u9032\u884c\u64f4\u589e\u53cd\u61c9\u3002<\/p>\n<p>\u3000\u3000\uff081\uff09\u5be6\u9a57\u524d\u914d\u5236\u9810\u6df7\u53cd\u61c9\u8a66\u5291\u4e26\u5206\u88dd\u3002\u9810\u6df7\u53cd\u61c9\u8a66\u5291\u5305\u62ec\u9664\u6a19\u672cDNA\u5916\u7684\u5176\u5b83\u5404\u7a2ePCR\u53cd\u61c9\u8a66\u5291\u3002\u6bcf\u652f\u53cd\u61c9\u7ba1\u4e2d\u542b\u6709\u7684PCR\u53cd\u61c9\u8a66\u5291\u898b\u88683\u3002<\/p>\n<p>\u3000\u3000\uff082\uff09\u5404\u53cd\u61c9\u7ba1\u4f9d\u6b21\u52a0\u516510&mu;l\u6a19\u672c\u548c90&mu;l\u9810\u6df7\u53cd\u61c9\u8a66\u5291\u3002<\/p>\n<p>\u3000\u3000\uff083\uff09\u52a0\u516580-100&mu;l\u77f3\u881f\u6cb9\uff0c\u5728\u81fa\u5f0f\u96e2\u5fc3\u6a5f\u4e0a\u5feb\u901f\u96e2\u5fc3\u6578\u79d2\u9418\uff0c\u4f7f\u5404\u53cd\u61c9\u8a66\u5291\u6536\u96c6\u65bc\u6cb9\u5c64\u4e0b\u3002\u76ee\u524d\uff0cPCR\u8a66\u5291\u5df2\u5546\u54c1\u5316\uff0c\u53cd\u61c9\u9ad4\u7a4d\u70ba25&mu;l\u3002\u4f7f\u7528\u6642\u53ea\u52a0\u5165\u6a19\u672cDNA\u5373\u53ef\u3002<\/p>\n<p>\u3000\u3000\uff084\uff09\u5c07\u53cd\u61c9\u7ba1\u7f6ePCR\u64f4\u589e\u5100\u4e0a\uff0c\u5faa\u74b0\u53c3\u6578\u70ba95\u2103 30s\uff0c55\u2103 40s\uff0c72\u2103 50s\u5faa\u74b035\u6b21\uff0c\u6700\u5f8c72\u2103\u5ef6\u4f385min\u3002 \u3000\u30004\uff0e\u6bcf\u6b21\u8a66\u9a57\u61c9\u8a2d\u967d\u6027\u53ca\u9670\u6027\u5c0d\u7167\u3002\u4ee5\u8f09\u6709HPV\u7684\u91cd\u7d44\u8cea\u7c92\uff08\u6bcf\u53cd\u61c9\u70ba100pg\uff09\u6216\u542b\u6709HPV\u7684\u7d30\u80de\u7cfb\uff08\u5982Caski\u3001HeLa\uff09DNA\u70ba\u967d\u6027\u5c0d\u7167\uff0c\u4ee5\u7121HPV\u7684\u4eba\u7d30\u80de\u7cfbDNA\u70ba\u9670\u6027\u5c0d\u7167\u3002<br \/>\n\u3000\u3000<br \/>\n\u3000\u3000\uff08\u4e09\uff09\u64f4\u589e\u7522\u7269\u7684\u6aa2\u6e2c\u548c\u5206\u6790<\/p>\n<p>\u3000\u30001\uff0e 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\/>\n\u3000\u3000\u7528PCR\u65b9\u6cd5\u6aa2\u6e2cHPV\u6bd4\u6838\u9178\u96dc\u4ea4\u65b9\u6cd5\u512a\u8d8a\u3002\u5176\u654f\u611f\u6027\u9ad8\uff0cGP-PCR\u65b9\u6cd5\u4ee5\u51dd\u81a0\u96fb\u6cf3\u76f4\u63a5\u5206\u6790\u7d50\u679c\uff0c\u53ef\u6aa2\u51fa\u6a19\u672c\u4e2d200\u500b\u62f7\u8c9d\u7684HPV DNA\uff0c\u82e5\u4ee5\u6838\u9178\u96dc\u4ea4\u6aa2\u6e2cPCR\u7522\u7269\uff0c\u654f\u611f\u6027\u63d0\u9ad8\uff0c\u80fd\u6aa2\u51fa10\u500b\u62f7\u8c9d\u7684HPV 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