CN101921748B - 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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- CN101921748B CN101921748B CN2010102137369A CN201010213736A CN101921748B CN 101921748 B CN101921748 B CN 101921748B CN 2010102137369 A CN2010102137369 A CN 2010102137369A CN 201010213736 A CN201010213736 A CN 201010213736A CN 101921748 B CN101921748 B CN 101921748B
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Abstract
Description
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(Mg2+ 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 |
Claims (40)
Priority Applications (3)
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CN2010102137369A CN101921748B (zh) | 2010-06-30 | 2010-06-30 | 用于高通量检测人类乳头瘤病毒的dna分子标签 |
PCT/CN2010/001812 WO2012000149A1 (zh) | 2010-06-30 | 2010-11-12 | 用于高通量检测人类乳头瘤病毒的dna分子标签 |
HK11103419.4A HK1149292A1 (en) | 2010-06-30 | 2011-04-05 | Dna molecular indexes for high-throughput detecting human papilloma virus |
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CN2010102137369A CN101921748B (zh) | 2010-06-30 | 2010-06-30 | 用于高通量检测人类乳头瘤病毒的dna分子标签 |
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CN101921874B (zh) | 2010-06-30 | 2013-09-11 | 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 |
CN102690809B (zh) * | 2011-03-24 | 2013-12-04 | 深圳华大基因科技服务有限公司 | Dna标签及其在构建和测序配对末端标签文库中的应用 |
CN102978206A (zh) * | 2012-11-27 | 2013-03-20 | 北京诺禾致源生物信息科技有限公司 | 一种应用于混合建库的高通量测序接头及其建库方法 |
WO2016050111A1 (en) * | 2014-09-30 | 2016-04-07 | Bgi Shenzhen | Biomarkers for rheumatoid arthritis and usage thereof |
CN105524983B (zh) * | 2014-09-30 | 2019-06-21 | 大连晶泰生物技术有限公司 | 基于高通量测序的标记和捕获多个样本的一个或多个特定基因的方法和试剂盒 |
WO2016049927A1 (en) * | 2014-09-30 | 2016-04-07 | Bgi Shenzhen Co., Limited | Biomarkers for obesity related diseases |
WO2016049918A1 (en) * | 2014-09-30 | 2016-04-07 | Bgi Shenzhen Co., Limited | Biomarkers for coronary artery disease |
WO2016049937A1 (en) * | 2014-09-30 | 2016-04-07 | Bgi Shenzhen Co., Limited | Biomarkers for rheumatoid arthritis and usage therof |
WO2016049930A1 (en) * | 2014-09-30 | 2016-04-07 | Bgi Shenzhen Co., Limited | Biomarkers for rheumatoid arthritis and usage therof |
CN109136217B (zh) * | 2017-06-27 | 2022-02-22 | 深圳华大基因股份有限公司 | 一种测序文库构建的方法、建库试剂及其应用 |
CN108130323A (zh) * | 2017-12-20 | 2018-06-08 | 奥明(杭州)基因科技有限公司 | 一种适用于植物转座酶可接近性染色质分析的建库方法 |
CN114410773B (zh) * | 2022-01-27 | 2024-05-03 | 宁波大学 | 用于预测或诊断抑郁症复发的标志物组合及其应用 |
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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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CN101250593B (zh) * | 2008-02-03 | 2011-01-26 | 广州益善生物技术有限公司 | 乳头瘤病毒检测与分型方法及其液相芯片 |
CN101435002B (zh) * | 2008-12-12 | 2012-01-18 | 深圳华大基因科技有限公司 | 一种检测人类乳头瘤病毒基因型的方法 |
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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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