CN103080336A - 检测胚胎或肿瘤染色体拷贝数的试剂盒、装置和方法 - Google Patents
检测胚胎或肿瘤染色体拷贝数的试剂盒、装置和方法 Download PDFInfo
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Abstract
Description
Claims (32)
- 权 利 要 求 书1. 一种检测胚胎或肿瘤染色体拷贝数的试剂盒, 包括:从母体或肿瘤患者体内取血液的器械;适合将所述血液中的血细胞和血浆分离的器械;抽提所述血浆中的 DNA的试剂和器械;通过物理方法根据所述 DNA 的片段大小对其进行分离的试剂和 器械; 和取 100bp-250bp之间的任意一点或任意一个区间的所有 DNA进行 测序的试剂和器械。
- 2. 根据权利要求 1所述的试剂盒, 其中所述胚胎或肿瘤染色体是整条染色 体或局部染色体。
- 3. 根据权利要求 1所述的试剂盒, 还包括: 将所述 DNA制成供测序的文 库的试剂和器械。
- 4. 根据权利要求 2所述的试剂盒, 还包括: 将所述 DNA制成供测序的文 库的试剂和器械。
- 5. 根据权利要求 4所述的试剂盒, 还包括: 对所述供测序的文库进行 PCR 扩增的试剂和器械。
- 6. 根据权利要求 1或 5所述的试剂盒, 其中所述 100-250bp之间的任意一 点或任意一个区间的 DNA是 150bp-170bp的 DNA。
- 7. 根据权利要求 6所述的试剂盒, 其中所述 100-250bp之间的任意一点或 任意一个区间的 DNA是 167bp的 DNA。
- 8. —种检测胚胎或肿瘤染色体拷贝数的试剂盒, 包括:从母体或肿瘤患者体内取血液的器械;适合将所述血液中的血细胞和血浆分离的器械;抽提所述血浆中的 DNA的试剂和器械;将所述 DNA制成可供测序的文库的试剂和器戈; 和对所述 DNA进行测序的试剂和器械。. 根据权利要求 8所述的试剂盒, 其中所述胚胎或肿瘤染色体是整条染色 体或局部染色体。
- 10. 根据权利要求 8 所述的试剂盒, 还包括: 对从血浆中抽提的所述 DNA 进行 PCR扩增的试剂和器械。
- 11. 一种检测胚胎或肿瘤染色体拷贝数的装置, 包括:检测模块,用于对母体血浆或肿瘤患者血浆样本中的 DNA进行测 序;比对模块,用于将所述 DNA的测序结果与染色体组序列图谱进行 比对, 以确定所述 DNA中的每段 DNA来自几号染色体以及所述每段 DNA的序列长度;计算模块,用于计算在同一样本内来自待测染色体的 DNA片段的 数量占 DNA片段总数的比值, 根据所述来自待测染色体的 DNA片段 中的 GC含量对所述比值进行校正; 并计算待测样本中所述来自待测 染色体的 DNA片段校正后的比值的变异,根据所述变异的程度确定待 测染色体的拷贝数; 以及输出模块, 用于输出所述待测染色体的拷贝数。
- 12. 根据权利要求 11所述的装置,其中所述计算模块根据以下的函数对第 2、 3、 4、 5、 6、 7、 8、 12、 13、 18 和 X号染色体的比值进行校正: 来自第 2、 3、 4、 5、 6、 7、 8、 12、 13、 18和 X号染色体的 DNA 片段的 GC含量分别与来自各个染色体的 DNA片段占总 DNA片段的比 值具有一定的线性关系, 所述线性关系可用 y=ax+b表示, 其中 y代表 来自待测染色体的 DNA片段的 GC含量, X代表来自待测染色体的 DNA 片段数量占总 DNA的比值, a和 b是常数, 且 a是负数。
- 13. 根据权利要求 12所述的装置, 其中所述来自第 2、 3、 4、 5、 6、 7、 8、 12、 13、 18和 X号染色体的 DNA片段的 GC含量与来自各个染色 体的 DNA片段占总 DNA片段的比值之间的线性关系如以下所示染色体编号 函数2 Y = -1274.5X + 154.483 Υ = -682.68Χ + 88.114 Y = -391.42X + 62.0755 Υ = -772.25Χ + 88.5176 Υ = -729.61X + 84.3547 Υ = -2874.7Χ + 197.288 Υ = -1599.4X + 122.8412 Υ = -1936.7X + 129.32 Y = -827.29X + 67.049Y = - 1946.4X + 97.323Υ = -749.77Χ + 71.33
- 14. 根据权利要求 11所述的装置,其中所述计算模块根据以下的函数对第 1、 9、 10、 11、 15、 16、 17、 19、 20、 21和 22号染色体的比值进行校正: 来自第 1、 9、 10、 11、 15、 16、 17、 19、 20、 21和 22号染色体的 DNA 片段的 GC含量分别与来自各个染色体的 DNA片段占总 DNA片段的比 值具有一定的线性关系, 所述线性关系可用 y=ax+b表示, 其中 y代表 来自待测染色体的 DNA片段的 GC含量, X代表来自待测染色体的 DNA 片段数量占总 DNA的比值, a和 b是常数, 且 a是正数。
- 15. 才艮据权利要求 14所述的装置, 其中所述来自第 1、 9、 10、 11、 15、 16、 17、 19、 20、 21和 22号染色体的 DNA片段的 GC含量与来自各个染色 体的 DNA片段占总 DNA片段的比值之间的线性关系如在以下所示: 染色体编号 函数1 Y = 909. 17X - 29.3729 Υ = 2211.3X - 45.63210 Υ = 1886.8X - 5 1.02611 Υ = 1255X - 15.71415 Υ = 1775.8Χ - 10. 12416 Υ = 797. 17X + 23.88317 Υ = 560.83Χ + 31.22719 Υ = 513.79X + 43.08820 Υ = 1006.7X + 22.81821 Υ = 7298Χ - 5 1.08322 Υ = 596.24Χ + 43.346
- 16. 根据权利要求 11- 15所述的装置, 其中所述检测模块将母体血浆或肿瘤 患者血浆样本中的所有 DNA制成可供测序的文库的过程。
- 17. 居权利要求 11所述的装置,其中所述对母体血浆或肿瘤患者血浆样本 中的 DNA进行测序是双端短序列测序、 单端长序列测序或单端短序列 测序。
- 18. 根据权利要求 11所述的装置,其中所述胚胎或肿瘤染色体是整条染色体 或局部染色体。
- 19. 根据权利要求 17所述的装置, 其中对所述母体血浆或肿瘤患者血浆样本 中的 DNA在测序前进行了 PCR扩增。
- 20. 根据权利要求 19 所述的装置, 其中计算模块用于计算在同一样本内 100bp-250bp之间的任意一点或任意一个区间的所有 DNA中来自待测染 色体的 DNA片段数量占 100bp-250bp之间的任意一点或任意一个区间 的所有 DNA片段总数的比值, 根据 100bp-250bp之间的任意一点或任 意一个区间的所有 DNA中来自待测染色体的 DNA片段的 GC含量对所 述比值进行校正; 并计算待测样本中所述来自待测染色体的 DNA 片段 校正后的比值的变异, 根据所述变异的程度确定待测染色体的拷贝数。
- 21. 居权利要求 20 所述的装置, 其中将所述母体血浆或肿瘤患者血浆样 本中 100bp-250bp之间的任意一点或任意一个区间的所有 DNA制成可 供测序的文库。
- 22. 居权利要求 21 所述的装置, 其中所述母体血浆或肿瘤患者血浆样本 中的 DNA在被制作成可供测序的文库之前或之后进行了 PCR扩增。
- 23. 根据权利要求 20所述的装置,其中所述 100-250bp之间的任意一点或任 意一个区间的 DNA是 150bp-170bp的 DNA。
- 24. 根据权利要求 23所述的装置,其中所述 100-250bp之间的任意一点或任 意一个区间是 DNA是 167bp的 DNA。
- 25. 一种检测胚胎或肿瘤染色体拷贝数的方法, 包括以下步骤:取母体血浆或肿瘤患者血浆;将所述血液中的血浆和血细月包分离;将所述血浆中的 DNA制作成可供测序的文库;对所述 DNA测序文库进行测序;将所述测序的结果与染色体组序列图谱进行比对, 确定每段所述 DNA序列来自几号染色体以及每段所述 DNA序列的长度; 以及用所述 DNA的测序和比对结果,计算在同一样本内来自待测染色 体的 DNA片段的数量占 DNA片段总数的比值, 根据所述来自待测染 色体的 DNA片段中的 GC含量对所述比值进行校正,并计算待测样本 中所述来自待测染色体的 DNA片段校正后的比值的变异,根据所述变 异的程度确定待测染色体的拷贝数。
- 26. 根据权利要求 25中任一项所述的方法, 根据以下的函数对第 2、 3、 4、 5、 6、 7、 8、 12、 13、 18和 X号染色体的比值进行校正: 来自第 2、 3、 4、 5、 6、 7、 8、 12、 13、 18和 X号染色体的 DNA片段的 GC 含量分别与来自各个染色体的 DNA片段占总 DNA片段的比值具有一定 的线性关系, 所述线性关系可用 y = ax + b表示, 其中 y代表来自待测 染色体的 DNA片段的 GC含量, X代表来自待测染色体的 DNA片段数 量占总 DNA的比值, a和 b是常数, 且 a是负数。
- 27. 根据权利要求 26所述的方法, 其中所述来自第 2、 3、 4、 5、 6、 7、 8、 12、 13、 18和 X号染色体的 DNA片段的 GC含量与来自各个染色 体的 DNA片段占总 DNA片段的比值之间的线性关系如在权利要求 13 中所示。
- 28. 根据权利要求 25中任一项所述的方法, 根据以下的函数对第 1、 9、 10、 11、 15、 16、 17、 19、 20、 21和 22号染色体的比值进行校正: 来自第 1、 9、 10、 11、 15、 16、 17、 19、 20、 21和 22号染色体的 DNA片段的 GC含量分别与来自各个染色体的 DNA片段占总 DNA片段的比值具有 一定的线性关系, 所述线性关系可用 y=ax+b表示, 其中 y代表来自待 测染色体的 DNA片段的 GC含量, X代表来自待测染色体的 DNA片段 数量占总 DNA的比值, a和 b是常数, JL a是正数。
- 29. 根据权利要求 28所述的方法, 其中所述来自第 1、 9、 10、 11、 15、 16、 17、 19、 20、 21和 22号染色体的 DNA片段的 GC含量与来自各个染色 体的 DNA片段占总 DNA片段的比值之间的线性关系如在权利要求 15 中所示。
- 30. 根据权利要求 25-29 所述的方法, 其中仅取 DNA 的序列长度在 100bp-250bp之间的任意一点或任意一个区间的所有 DNA的测序和比对 结果, 计算在同一样本内 100bp-250bp之间的任意一点或任意一个区间 的所有 DNA中来自待测染色体的 DNA片段数量占 100bp-250bp之间的 任意一点或任意一个区间的所有 DNA 片段总数的比值, 根据 100bp-250bp之间的任意一点或任意一个区间的所有 DNA中所述来自待 测染色体的 DNA片段中的 GC含量对所述比值进行校正, 并计算待测 样本中所述来自待测染色体的 DNA 片段校正后的比值的变异, 根据所 述变异的程度确定待测染色体的拷贝数。
- 31. 根据权利要求 25-29所述的方法, 其中对所述 DNA测序文库进行测序 是双端短序列测序、 单端长序列测序或单端短序列测序。
- 32. 根据权利要求 30所述的方法,其中所述 100-250bp之间的任意一点或任 意一个区间的 DNA是 150bp-170bp的 DNA。
- 33. 根据权利要求 32所述的方法,其中所述 100-250bp之间的任意一点或任 意一个区间的 DNA是 167bp的 DNA。
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