CN101921748A - 用于高通量检测人类乳头瘤病毒的dna分子标签 - Google Patents
用于高通量检测人类乳头瘤病毒的dna分子标签 Download PDFInfo
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- CN101921748A CN101921748A CN 201010213736 CN201010213736A CN101921748A CN 101921748 A CN101921748 A CN 101921748A CN 201010213736 CN201010213736 CN 201010213736 CN 201010213736 A CN201010213736 A CN 201010213736A CN 101921748 A CN101921748 A CN 101921748A
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  标签编号 |   标签序列 |   对应96孔板位置 |  对应样品(组1) |  对应样品(组2) |  SEQ ID NO: |
  PI-1 |   GCTGCGACTC |   A1 |  1 |  96 |  1 |
  PI-2 |   GTGTAGATAC |   A2 |  2 |  97 |  2 |
  PI-3 |   CTGATATCTA |   A3 |  3 |  98 |  3 |
  PI-4 |   ACGATGCTAT |   A4 |  4 |  99 |  4 |
  PI-5 |   TAGACTAGAC |   A5 |  5 |  100 |  5 |
  PI-6 |   CTGTCTGTGT |   A6 |  6 |  101 |  6 |
  PI-7 |   GCATACTGAC |   A7 |  7 |  102 |  7 |
  PI-8 |   CTGCTCGCAT |   A8 |  8 |  103 |  8 |
  PI-9 |   CATGAGTAGA |   A9 |  9 |  104 |  9 |
  PI-10 |   TCTCACTATG |   A10 |  10 |  105 |  10 |
  PI-11 |   TGTACTACTA |   A11 |  11 |  106 |  11 |
  PI-12 |   GTAGACTAGT |   A12 |  12 |  107 |  12 |
  PI-13 |   ATATGCTACT |   B1 |  13 |  108 |  13 |
  PI-14 |   CACTCGCTGT |   B2 |  14 |  109 |  14 |
  PI-15 |   CATCACGCAC |   B3 |  15 |  110 |  15 |
  PI-16 |   AGCATGTGAT |   B4 |  16 |  111 |  16 |
  PI-17 |   AGCTAGTAGA |   B5 |  17 |  112 |  17 |
  PI-18 |   GCTATGTAGT |   B6 |  18 |  113 |  18 |
  PI-19 |   TACGATGATG |   B7 |  19 |  114 |  19 |
  PI-20 |   TACGCTGTAC |   B8 |   20 |   115 |   20 |
  PI-21 |   TATGTGTACT |   B9 |   21 |   116 |   21 |
  PI-22 |   TGACTCAGAC |   B10 |   22 |   117 |   22 |
  PI-23 |   TCGTAGCTCA |   B11 |   23 |   118 |   23 |
  PI-24 |   GAGACTCGTA |   B12 |   24 |   119 |   24 |
  PI-25 |   CTAGATGTCA |   C1 |   25 |   120 |   25 |
  PI-26 |   GATGACTCTC |   C2 |   26 |   121 |   26 |
  PI-27 |   TCAGTCGCAC |   C3 |   27 |   122 |   27 |
  PI-28 |   TGTAGTGAGT |   C4 |   28 |   123 |   28 |
  PI-29 |   TCATCGTAGA |   C5 |   29 |   124 |   29 |
  PI-30 |   TAGCATCTGT |   C6 |   30 |   125 |   30 |
  PI-31 |   TAGTAGTCGT |   C7 |   31 |   126 |   31 |
  PI-32 |   CTATACGTGC |   C8 |   32 |   127 |   32 |
  PI-33 |   CGACTGTAGA |   C9 |   33 |   128 |   33 |
  PI-34 |   GATGTCATGT |   C10 |   34 |   129 |   34 |
  PI-35 |   GTCTCGACTG |   C11 |   35 |   130 |   35 |
  PI-36 |   AGCTGACGAT |   C12 |   36 |   131 |   36 |
  PI-37 |   ATGATATAGT |   D1 |   37 |   132 |   37 |
  PI-38 |   ATGTGCTCTA |   D2 |   38 |   133 |   38 |
  PI-39 |   CTCACTCGAT |   D3 |   39 |   134 |   39 |
  PI-40 |   GCTGCGACTC |   D4 |   40 |   135 |   40 |
  PI-41 |   GAGTCATGTC |   D5 |   41 |   136 |   41 |
  PI-42 |   CATACGCTCA |   D6 |   42 |   137 |   42 |
  PI-43 |   CACTCTCGTC |   D7 |   43 |   138 |   43 |
  PI-44 |   GCACTAGATG |   D8 |   44 |   139 |   44 |
  PI-45 |   AGTACGCATG |   D9 |   45 |   140 |   45 |
  PI-46 |   TCTGTGACGT |   D10 |   46 |   141 |   46 |
  PI-47 |   TAGCTCATCT |   D11 |   47 |   142 |   47 |
  PI-48 |   AGCATACACT |   D12 |   48 |   143 |   48 |
  PI-49 |   GCTATAGTCA |   E1 |   49 |   144 |   49 |
  PI-50 |   CGTCTCATGC |   E2 |   50 |   145 |   50 |
  PI-51 |   GCTACTACGT |   E3 |   51 |   146 |   51 |
  PI-52 |   GAGTGTACTA |   E4 |   52 |   147 |   52 |
  PI-53 |   GTCATACGTG |   E5 |   53 |   148 |   53 |
  PI-54 |   TATGAGAGAT |   E6 |   54 |   149 |   54 |
  PI-55 |   ATCTGAGTAC |   E7 |   55 |   150 |   55 |
  PI-56 |   CGATAGCATC |   E8 |   56 |   151 |   56 |
  PI-57 |   ACTGATCTCA |   E9 |   57 |   152 |   57 |
  PI-58 |   CTCGATACTA |   E10 |   58 |   153 |   58 |
  PI-59 |   CATGTGACTG |   E11 |   59 |   154 |   59 |
  PI-60 |   CGCATCACTA |   E12 |   60 |   155 |   60 |
  PI-61 |   GCATATATCT |   F1 |   61 |   156 |   61 |
  PI-62 |   CTGATGCGAC |   F2 |   62 |   157 |   62 |
  PI-63 |   TCTCAGAGTC |   F3 |   63 |   158 |   63 |
  PI-64 |   CAGTGCGAGT |   F4 |   64 |   159 |   64 |
  PI-65 |   ATCTCTGATG |   F5 |   65 |   160 |   65 |
  PI-66 |   GCTAGTAGTC |   F6 |   66 |   161 |   66 |
  PI-67 |   ATGAGTCGTC |   F7 |   67 |   162 |   67 |
  PI-68 |   ATCACTCAGA |   F8 |   68 |   163 |   68 |
  PI-69 |   TCTCTCTGAT |   F9 |   69 |   164 |   69 |
  PI-70 |   CTCTAGTGCT |   F10 |   70 |   165 |   70 |
  PI-71 |   CGTCGTGCTA |   F11 |   71 |   166 |   71 |
  PI-72 |   CGACTACTAT |   F12 |   72 |   167 |   72 |
  PI-73 |   GCACGTCGAT |   G1 |   73 |   168 |   73 |
  PI-74 |   GTAGTGCTCT |   G2 |   74 |   169 |   74 |
  PI-75 |   CTGACGAGCT |   G3 |   75 |   170 |   75 |
  PI-76 |   CTATAGTCTA |   G4 |   76 |   171 |   76 |
  PI-77 |   ACACGCACTA |   G5 |   77 |   172 |   77 |
  PI-78 |   CTCGCACTAC |   G6 |   78 |   173 |   78 |
  PI-79 |   AGATCTCACT |   G7 |   79 |   174 |   79 |
  PI-80 |   ATACTAGTGT |   G8 |   80 |   175 |   80 |
  PI-81 |   ATATCTCGTA |   G9 |   81 |   176 |   81 |
  PI-82 |   TGACTGCGTA |   G10 |   82 |   177 |   82 |
  PI-83 |   TGTAGACGTA |   G11 |   83 |   178 |   83 |
  PI-84 |   AGAGACTATG |   G12 |   84 |   179 |   84 |
  PI-85 |   GTCGAGTCAC |   H1 |   85 |   180 |   85 |
  PI-86 |   TGACAGCTAC |   H2 |   86 |   181 |   86 |
  PI-87 |   CGCTAGACAT |   H3 |   87 |   182 |   87 |
  PI-88 |   CGTAGATATG |   H4 |   88 |   183 |   88 |
  PI-89 |   TGAGTCTGCT |   H5 |   89 |   184 |   89 |
  PI-90 |   TAGTCGTATG |   H6 |   90 |   185 |   90 |
  PI-91 |   CATACACGAC |   H7 |   91 |   186 |   91 |
  PI-92 |   CGCTCAGAGA |   H8 |   92 |   187 |   92 |
  PI-93 |   GTGAGTCTCA |   H9 |   93 |   188 |   93 |
  PI-94 |   GACAGATGAT |   H10 |   94 |   189 |   94 |
  PI-95 |   GCTGTGCGAC |   H11 |   95 |   190 |   95 |
  引物编号 |   引物序列 |   SEQ ID NO: |
  F1 |   TTTGTTACTGTGGTAGATACTAC |   96 |
  F2 |   TTTGTTACTGTGGTGGATACTAC |   97 |
  F3 |   TTTGTTACCGTTGTTGATACTAC |   98 |
  F4 |   TTTGTTACTAAGGTAGATACCACTC |   99 |
  F5 |   TTTGTTACTGTTGTGGATACAAC |   100 |
  F6 |   TTTGTTACTATGGTAGATACCACAC |   101 |
  R1 |   GAAAAATAAACTGTAAATCATATTCCT |   102 |
  R2 |   GAAAAATAAATTGTAAATCATACTC |   103 |
  R3 |   GAAATATAAATTGTAAATCAAATTC |   104 |
  R4 |   GAAAAATAAACTGTAAATCATATTC |   105 |
  R5 |   GAAAAATAAACTGCAAATCATATTC |   106 |
  试剂 |   体积/反应 |
  水(HPLC级) |   14.375μl |
  10x Ex Taq缓冲液(Mg<sup>2+</sup> plus) |   2.5μl |
  dNTP混合物(各2.5mM) |   2μl |
  带有标签的F1/F2/F3/F4/F5/F6的混合物(各7.5pmol) |   0.5μl |
  带有标签的R1/R2/R3/R4/R5的混合物(各7.5pmol) |   0.5μl |
  Ex Taq HS(5U/μl) |   0.125μl |
  模板DNA |   5μl |
  总体积 |   25μl |
  试剂 |   体积/反应 |
  上一步所得的DNA |   75μl |
  20x多核苷酸激酶缓冲液(B904) |   10μL |
  dNTP混合物(各20mM) |   4μL |
  T4 DNA聚合酶 |   5μL |
  Klenow片段 |   1μL |
  T4多核苷酸激酶 |   5μL |
  总体积 |   100μL |
  试剂 |   体积/反应 |
  上一步所得的DNA |   32μl |
  dATP(1mM,GE公司) |   10μl |
  10x Blue缓冲液 |   5μl |
  Klenow (3′-5′exo-) |   3μl |
  总体积 |   50μl |
  试剂 |   体积/反应 |
  上一步所得的DNA |   11μL |
  2x快速连接缓冲液 |   15μL |
  PCR-free Index Adapter Oligo Mix(25mM) |   1μL |
  T4DNA连接酶(快速,L603-HC-L) |   3μL |
  总体积 |   30μL |
  2100(nM) |   qPCR(nM) | |
  HPV-1组 |   20.4 |   24.2 |
  HPV-2组 |   21.6 |   25.8 |
  样品  编号 |   原HC-II结果  (RLU/CO值) |   本次检测的  HPV结果 |   样品  编号 |   原HC-II结果  (RLU/CO值) |   本次检测的  HPV结果 |
  1 |   14.2 |   HPV56 |   96 |   0.28 |   阴性 |
  2 |   0.31 |   阴性 |   97 |   0.33 |   阴性 |
  3 |   196.41 |   HPV16 |   98 |   181.29 |   HPV35,HPV6 |
  4 |   5.76 |   HPV18 |   99 |   77.32 |   HPV16 |
  5 |   0.35 |   阴性 |   100 |   91.22 |   HPV39 |
  6 |   99.86 |   HPV18,HPV11,HPV16 |   1011 |   188.92 |   HPV52 |
  7 |   128.86 |   HPV39 |   102 |   1352.83 |   HPV35,HPV11,HPV39 |
  8 |   35.12 |   HPV18,HPV6 |   103 |   1.39 |   HPV43 |
  9 |   498.69 |   HPV16,HPV56 |   104 |   119.5 |   HPV45,HPV11 |
  10 |   603.57 |   HPV18,HPV31,HPV39 |   105 |   292.43 |   HPV56,HPV31 |
  11 |   0.27 |   阴性 |   106 |   2.91 |   HPV68 |
  12 |   3420.57 |   HPV18 |   107 |   193.13 |   HPV45 |
  13 |   0.38 |   阴性 |   108 |   2.62 |   HPV6 |
  14 |   0.41 |   阴性 |   109 |   94.12 |   HPV16 |
  15 |   455.06 |   HPV16 |   110 |   792.72 |   HPV18,HPV31 |
  16 |   8.93 |   HPV18 |   111 |   31.76 |   HPV11 |
  17 |   0.6 |   阴性 |   112 |   0.25 |   阴性 |
  18 |   0.41 |   阴性 |   113 |   0.23 |   阴性 |
  19 |   0.29 |   阴性 |   114 |   750.82 |   HPV56,HPV16 |
  20 |   27.64 |   HPV31 |   115 |   0.4 |   阴性 |
  21 |   1985.41 |   HPV56,HPV68 |   116 |   2.75 |   HPV31 |
  22 |   20.71 |   HPV42 |   117 |   396.04 |   HPV45 |
  23 |   1795.83 |   HPV11,HPV16,HPV52 |   118 |   354.76 |   HPV18,HPV16 |
  24 |   9.55 |   HPV43 |   119 |   6.26 |   HPV11 |
  25 |   237.62 |   HPV39 |   120 |   1719.67 |   HPV16,HPV45 |
  26 |   1.5 |   HPV6 |   121 |   76.92 |   HPV51 |
  27 |   1478.98 |   HPV68,HPV16 |   122 |   1318.02 |   HPV56,HPV16,HPV42 |
  28 |   115.31 |   HPV44 |   123 |   0.28 |   阴性 |
  29 |   419.31 |   HPV16 |   124 |   0.33 |   阴性 |
  30 |   1.81 |   candHPV89 |   125 |   181.29 |   HPV59 |
  31 |   2013.61 |   HPV52,HPV39 |   126 |   77.32 |   HPV68 |
  32 |   1379.09 |   HPV54,HPV33 |   127 |   110.8 |   HPV52 |
  33 |   12.74 |   HPV42 |   128 |   147.25 |   HPV16 |
  34 |   1695.31 |   HPV16,candHPV89 |   129 |   0.24 |   HPV26 |
  35 |   1410.85 |   HPV35 |   130 |   1.55 |   HPV11,HPV53 |
  36 |   1149.25 |   HPV18 |   131 |   2.03 |   HPV6,HPV66 |
  37 |   0.24 |   阴性 |   132 |   8.45 |   HPV43 |
  38 |   1.55 |   HPV11 |   133 |   0.2 |   阴性 |
  39 |   2.03 |   HPV11,HPV6 |   134 |   0.24 |   阴性 |
  40 |   8.45 |   HPV42 |   135 |   10.53 |   HPV11 |
  41 |   0.2 |   阴性 |   136 |   1410.85 |   HPV16,HPV53,HPV70 |
  42 |   0.22 |   阴性 |   137 |   1149.25 |   HPV56,HPV81,HPV73 |
  43 |   0.53 |   阴性 |   138 |   0.24 |   阴性 |
  44 |   10.38 |   HPV6 |   139 |   413.9 |   HPV45 |
  45 |   78.21 |   HPV16 |   140 |   17.05 |   HPV11 |
  46 |   0.23 |   阴性 |   141 |   23.6 |   HPV52 |
  47 |   45.42 |   HPV16,HPV18 |   142 |   3379.09 |   HPV16,HPV35,HPV56 |
  48 |   0.35 |   阴性 |   143 |   0.18 |   阴性 |
  49 |   148.66 |   HPV18,candHPV89 |   144 |   1.46 |   HPV18 |
  50 |   60.27 |   HPV56 |   145 |   1.25 |   HPV11,HPV26 |
  51 |   0.28 |   阴性 |   146 |   2.13 |   HPV6,HPV81 |
  52 |   360.26 |   HPV56,HPV68 |   147 |   872.52 |   HPV16,HPV45,HPV52 |
  53 |   50.31 |   HPV18 |   148 |   1.5 |   HPV18 |
  54 |   0.18 |   阴性 |   149 |   4.33 |   HPV16 |
  55 |   0.31 |   阴性 |   150 |   0.82 |   阴性 |
  56 |   196.41 |   HPV16 |   151 |   60.35 |   HPV59 |
  57 |   5.76 |   HPV51 |   152 |   0.24 |   阴性 |
  58 |   0.23 |   阴性 |   153 |   0.23 |   阴性 |
  59 |   0.88 |   阴性 |   154 |   0.18 |   阴性 |
  60 |   0.16 |   阴性 |   155 |   1.46 |   HPV51 |
  61 |   870.63 |   HPV52,HPV16 |   156 |   11.25 |   HPV16 |
  62 |   10.18 |   HPV42 |   157 |   2.13 |   HPV11 |
  63 |   0.15 |   阴性 |   158 |   0.13 |   阴性 |
  64 |   1.36 |   HPV11 |   159 |   90.18 |   HPV58 |
  65 |   68.2 |   HPV59 |   160 |   0.15 |   阴性 |
  66 |   0.68 |   阴性 |   161 |   602.79 |   HPV68,HPV16 |
  67 |   130.41 |   HPV45 |   162 |   132.68 |   HPV56,HPV11 |
  68 |   0.26 |   阴性 |   163 |   127.08 |   HPV39,HPV54 |
  69 |   5.25 |   HPV6 |   164 |   602.79 |   HPV33 |
  70 |   0.46 |   阴性 |   165 |   276 |   HPV18 |
  71 |   8.23 |   HPV40 |   166 |   243.6 |   HPV45 |
  72 |   0.28 |   阴性 |   167 |   229.44 |   HPV51 |
  73 |   100.16 |   HPV43,HPV44 |   168 |   1384.92 |   HPV16,HPV58,HPV72 |
  74 |   450.13 |   HPV41 |   169 |   172.64 |   HPV58 |
  75 |   127.08 |   HPV39,HPV6 |   170 |   855.24 |   HPV16,candHPV89 |
  76 |   602.79 |   HPV45 |   171 |   126.47 |   HPV51 |
  77 |   276 |   HPV16 |   172 |   86.62 |   HPV44,HPV11 |
  78 |   243.6 |   HPV6,HPV70,HPV39 |   173 |   879.37 |   HPV18,HPV58 |
  79 |   229.44 |   HPV35 |   174 |   119.39 |   HPV56 |
  80 |   1384.92 |   HPV52,HPV56,HPV11 |   175 |   0.61 |   阴性 |
  81 |   172.64 |   HPV26,HPV42 |   176 |   18.02 |   HPV16 |
  82 |   855.24 |   HPV35,HPV6 |   177 |   16.06 |   HPV18 |
  83 |   620.69 |   HPV52 |   178 |   60.69 |   HPV56,HPV11 |
  84 |   128.02 |   HPV11 |   179 |   2.45 |   HPV11 |
  85 |   514.84 |   HPV33 |   180 |   94.93 |   HPV39 |
  86 |   68.3 |   HPV58 |   181 |   1635.3 |   HPV16,HPV35,HPV51 |
  87 |   402.15 |   HPV59,HPV16 |   182 |   754.64 |   HPV33,candHPV89 |
  88 |   51.72 |   HPV33 |   183 |   0.23 |   HPV11 |
  89 |   1.78 |   HPV6 |   184 |   20.28 |   HPV18 |
  90 |   56.7 |   HPV11,HPV31 |   185 |   0.16 |   阴性 |
  91 |   186.06 |   HPV16 |   186 |   0.13 |   阴性 |
  92 |   0.02 |   阴性 |   187 |   60.18 |   HPV59 |
  93 |   386.06 |   HPV18,HPV16 |   188 |   0.15 |   阴性 |
  94 |   28.09 |   HPV6,HPV44 |   189 |   1.36 |   HPV43 |
  95 |   186.06 |   HPV68, |   190 |   0.28 |   阴性 |
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WO2012000151A1 (zh) * | 2010-06-30 | 2012-01-05 | 深圳华大基因科技有限公司 | 基于solexa测序法的检测人类乳头瘤病毒的方法 |
CN102690809A (zh) * | 2011-03-24 | 2012-09-26 | 深圳华大基因科技有限公司 | Dna标签及其在构建和测序配对末端标签文库中的应用 |
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CN101624632A (zh) * | 2008-07-08 | 2010-01-13 | 天津生物芯片技术有限责任公司 | 人乳头瘤病毒基因芯片、制备方法、应用及检测用试剂盒与应用 |
CN101921874A (zh) * | 2010-06-30 | 2010-12-22 | 深圳华大基因科技有限公司 | 基于Solexa测序法的检测人类乳头瘤病毒的方法 |
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