CN113528656B - 评价胶质瘤和/或胃腺癌预后性的试剂盒和系统 - Google Patents
评价胶质瘤和/或胃腺癌预后性的试剂盒和系统 Download PDFInfo
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Abstract
本公开涉及一种评价胶质瘤和/或胃腺癌预后性的试剂盒和系统，其中，该试剂盒包括能够特异性地扩增SEQ ID NO.1的cDNA片段的第一引物对，和/或，能够与SEQ ID NO.1的cDNA片段特异性杂交的第一探针，其中，所述SEQ ID NO:1的cDNA片段至少含有SEQ ID NO:1的第4057‑4074位、第4067‑4084位或者第4064‑4079位的cDNA序列。通过上述技术方案，本公开有效地提高了胶质瘤和/或胃腺癌预后性评价的准确性。
Description
技术领域
本公开涉及生物技术领域，具体地，涉及一种用于评价胶质瘤和/或胃腺癌预后性的试剂盒以及一种用于评价胶质瘤和/或胃腺癌预后性的系统。
背景技术
胶质瘤是源自神经上皮的肿瘤的统称，占颅脑肿瘤的40％-50％，是最常见的原发性颅内肿瘤。胶质瘤难以根治，手术后往往会复发，而且预后性不佳。世界卫生组织分级系统根据胶质瘤的组织学特征将胶质瘤分为Ⅰ-Ⅳ级，不同级别胶质瘤的预后性各不相同。
准确评价胶质瘤患者的预后性具有重要的临床、科研及社会价值。在临床工作中，准确的预后性评估可指导医生针对高风险患者制定个性化的检查及治疗方案，帮助医生制定合理的复查及随访计划，进而提高医疗服务的质量。在科研中，准确评估患者预后性的风险层级，可以为研发针对高风险病人的有效治疗方案提供重要依据，并且可以成为检验新型治疗效果的重要参考。从社会角度来说，准确评估患者预后性，可为病患及家属提供科学的生存预期，指引病人依从治疗计划，避免过度医疗，减轻家庭经济压力，有助于改善医患关系。
目前，主要依据胶质瘤的病理级别进行胶质瘤的预后性评价，但是同一病理级别的不同胶质瘤患者的预后性也会存在较大差异，仅依据胶质瘤病理级别进行胶质瘤的预后性评价仍然不够准确。
发明内容
本公开的目的是提供一种用于评价胶质瘤和/或胃腺癌预后性的试剂盒以及一种用于评价胶质瘤和/或胃腺癌预后性的系统，能够进一步提高胶质瘤和/或胃腺癌预后评价的准确性。
第一方面，本公开提供一种评价胶质瘤和/或胃腺癌预后性的试剂盒，该试剂盒包括能够特异性地扩增SEQ ID NO.1的cDNA片段的第一引物对，和/或，能够与SEQ ID NO.1的cDNA片段特异性杂交的第一探针，其中，所述SEQ ID NO:1的cDNA片段至少含有SEQ ID NO:1的第4057-4074位、第4067-4084位或者第4064-4079位的cDNA序列。
可选地，所述SEQ ID NO:1的cDNA片段至少含有SEQ ID NO:1的第4023-4122位的cDNA序列；优选地，所述SEQ ID NO:1的cDNA片段是指如SEQ ID NO:1所示的cDNA序列。
可选地，所述第一引物对含有如SEQ ID NO:4所示的第一引物和SEQ ID NO:5所示的第二引物，所述第一探针的序列中含有如SEQ ID NO:6所示的序列。
第二方面，本公开提供一种评价胶质瘤和/或胃腺癌预后性的试剂盒，该试剂盒包括能够特异性地扩增SEQ ID NO.2的mRNA片段的第二引物对，和/或，能够与SEQ ID NO.2的mRNA片段特异性杂交的第二探针，其中，所述SEQ ID NO:2的mRNA片段至少含有SEQ ID NO:2的第2452-2469位、第2464-2481位或者第2458-2474位的mRNA序列。
可选地，所述SEQ ID NO:2的mRNA片段至少含有SEQ ID NO:2的第2416-2516位的mRNA序列；优选地，所述SEQ ID NO:2的mRNA片段是指如SEQ ID NO:2所示的mRNA序列。
可选地，所述第二引物对含有如SEQ ID NO:7所示的第三引物和SEQ ID NO:8所示的第四引物，所述第二探针的序列中含有如SEQ ID NO:9所示的序列。
第三方面，本公开提供一种评价胶质瘤和/或胃腺癌预后性的试剂盒，该试剂盒包括抗SEQ ID NO.3的氨基酸片段的抗体，其中，所述SEQ ID NO.3的氨基酸片段至少含有SEQID NO:3的第750-764位的氨基酸序列。
可选地，所述SEQ ID NO.3的氨基酸片段至少含有SEQ ID NO:3的第722-764位的氨基酸序列；优选地，所述SEQ ID NO.3的氨基酸片段是指如SEQ ID NO:3所示的氨基酸序列。
第四方面，本公开提供一种分子试剂在制备评价胶质瘤和/或胃腺癌预后性的试剂盒中的用途，所述分子试剂包括如下(1)-(8)中的至少一种：
(1)如SEQ ID NO.1所示的cDNA；
(2)能够特异性地扩增如SEQ ID NO.1所示的cDNA的第一引物对；
(3)能够与如SEQ ID NO.1所示的cDNA特异性杂交的第一探针；
(4)如SEQ ID NO.2所示的mRNA；
(5)能够特异性地扩增如SEQ ID NO.2所示的mRNA的第二引物对；
(6)能够与如SEQ ID NO.2所示的mRNA特异性杂交的第二探针；
(7)如SEQ ID NO.3所示的蛋白；
(8)抗如SEQ ID NO.3所示的蛋白的抗体。
第五方面，本公开提供一种评价胶质瘤和/或胃腺癌预后性的系统，该系统包括扩增装置、测序装置、计算装置和输出装置；
所述扩增装置含有采集单元和扩增单元，所述采集单元用于采集模板核酸片段和扩增引物，所述扩增单元用于利用所述扩增引物对所述模板核酸片段进行扩增，得到扩增产物；
所述测序装置用于对所述扩增产物进行核酸序列测序，得到扩增产物序列；
所述计算装置包括存储器和处理器，所述存储器中存储有计算机程序，所述处理器被配置为执行所述存储器中存储的计算机程序，以实现如下判别：
若所述扩增产物序列中含有如SEQ ID NO.1所示的cDNA的序列和/或如SEQ IDNO.2所示的mRNA的序列，则判定所述模板核酸片段对应的胶质瘤和/或胃腺癌预后性较差；
所述输出装置用于输出所述计算装置的判定结果。
通过上述技术方案，本公开有效地提高了胶质瘤和/或胃腺癌预后性评价的准确性。
本公开的其他特征和优点将在随后的具体实施方式部分予以详细说明。
附图说明
附图是用来提供对本公开的进一步理解，并且构成说明书的一部分，与下面的具体实施方式一起用于解释本公开，但并不构成对本公开的限制。在附图中：
图1是本公开实施例提供的cDNA琼脂糖凝胶核酸电泳图；
图2是本公开实施例提供的cDNA Sanger测序的结果图；
图3是本公开实施例提供的全级别胶质瘤样本患者生存曲线图；
图4是本公开实施例提供的IDH突变的全级别胶质瘤样本患者生存曲线图；
图5是本公开实施例提供的继发胶质母细胞瘤样本患者生存曲线图；
图6是本公开实施例提供的IDH突变的继发胶质母细胞瘤样本患者生存曲线图；
图7是本公开实施例提供的WHO Ⅲ级的胃腺癌样本患者生存曲线图。
具体实施方式
以下结合附图对本公开的具体实施方式进行详细说明。应当理解的是，此处所描述的具体实施方式仅用于说明和解释本公开，并不用于限制本公开。
本公开的第一方面提供一种评价胶质瘤和/或胃腺癌预后性的试剂盒，该试剂盒包括能够特异性地扩增SEQ ID NO.1的cDNA片段的第一引物对，和/或，能够与SEQ ID NO.1的cDNA片段特异性杂交的第一探针，其中，所述SEQ ID NO:1的cDNA片段至少含有SEQ IDNO:1的第4057-4074位、第4067-4084位或者第4064-4079位的cDNA序列。
优选地，所述SEQ ID NO:1的cDNA片段至少含有SEQ ID NO:1的第4023-4122位的cDNA序列；优选地，所述SEQ ID NO:1的cDNA片段是指如SEQ ID NO:1所示的cDNA序列。
上述SEQ ID NO:1所示的cDNA序列是10号外显子缺失的人MET基因对应的cDNA，本公开的发明人发现，当胶质瘤和/或胃腺癌细胞的mRNA可逆转录出上述cDNA时，胶质瘤和/或胃腺癌的恶性程度更高，患者的预后性更差。其中，上述SEQ ID NO:1的cDNA片段中含有人MET基因10号外显子的缺失位点，SEQ ID NO:1所示的cDNA序列中含有人MET基因的除10号外显子外的所有外显子，SEQ ID NO:1所示的序列如下所示：
tacaagatgttgcatcactttactttaattgcatgatttatcagaacaactattaacatacgaagtaccattcagttcagctgcaggtataggcagtgacaagtatctaattcttagaagaatcacttactcccacaatctgtccagacacattaatctaaggacaagtttataaatagcaaacgtgattttcacattgcagtgttctcaagaatgtatatacaagtgtgtagtcctgttgatgggatgtttccccgagttctttctattgatgcgttcatgctcttgaccctggtagagacagttctttctttccacagagcagattttcttttgtcatccaccatttacaatactctgtgaattagtattacttattacatttatgaaacggcaatatttggattgtaatagctcttcagtacaattccttgtgtcttctggtaagtctctgcactatcagagagcttttagttataatcattttcctgcaacaacagccaaactcaactattgagtttcagtgtgacacaccctctttgtagcttgctgggttaacccttggcttcaagtcctgatgatgtaatgagggtggggtgtattggcaatcagtacatttccttatcgcaatttacagtcattgaaaatcatgctgtcattaatcccagtctgacataccttttctaaaatgttcacagtgcagtgtttttgtggcctaacaaaatttttctcatatcattaaaaataaacatttttataaaaaatataacactttaaatgtttacgtcgacaaaaccagttagagtaacctacaccacatgcactatacagtagcaagcacaaaattccacagaatgaagcatcacaaagttctgctcagggtggctattccatctaggtgaaatagctgggattttcaattgcctttttcatttgtttctaaagtatgttttgcttaacataaaacacaccctaatgcaaaataaaactccccaaaagttttgtttccaattgcttgcgaggtgggaacctgccaccgagacagaggctaatcttttcaatccatccaccctttctttgctctacctatgagctgtgattggaaccaatgaaccttttagtaaaatgtatcctgctttacaaacatgctgagttatctttaaaaatatttatcaacaaattacttgtcttattttgagttttcatttaaaaaaatacacacaaaacatctacatgttcacattcattagatcagagtagcatcattctcaaacagtgggtttttatatgacaactaaatatttcatgatgactaaattatttcatacaaattttcttaatgtcatctatgtacattacatcctgttcatttataaacatgttttgtgtgatgctattttgaaactttgactgtattgcctataaatattcttccaaccaaaggaatgtctatacaaaaatttataagggctgggcgcagtggctcatgcctgtaatcctagcactttgggaggctgaggtgggtggatttcttgagtccaggagtttgaaaccagccttggcaacatggcaaaaccccgtctctacaaaaaatacaaaaattagccggggatggtggtgtgcgcctgtagtcccagctattcgggaggcttgagcctgggaggtggaggttgcagtgagctatgatcacaccactgcactacagccctggcaacagagtgagatcctgtctcaaaaaacaaacaaacatttaaaagggaatgtttacctaatgggtgaatgacaccatcagtaatgaaaaatgttattgaccatacaacacacaaaagtgttttgaaacactagaattctaaatgagtggcctgttctggggctgccgctcctgtcctgagcattactatctcttctgagttttctgtgatcaagaagccctcaatatttcctgcttcaatttcccatatgaaatcaagtggtacctgatttttaaaaaattcaacccagaattcaagttttggactccaatatgacaggagtgttgtcacggctgcaggccattggtccgtggcctgtgggaccaagcctctggttctgatgctctgtcagataagaaattccttagaatccagtaatttaaataacaatacaagtcctataatagtgcaattttggcaagagcaaagaatatcgatggccttttaaaggtcaggcagtgaaaaaaccattggacaaagtgtggactgttgctttgacatagtactagcactatgatgtctcccagaaggaggctggtcgtgtgtccacctcatcatcagcgttatcttctgatgacaacagagaaggatacggagcgacacattttacgttcacataagtagcgttcacatggacatagtgctccccaatgaaagtagagaagatcgctgatatccgggacaccagttcagaaaaggatgggcgcatttcggctttagggtgccagcattttagcattacttcatataaggggtctgggcagtattcgggttgtaggagtcttctcccttgcaacaagtaaacagttatatcaaaggtgtttacgtcaggataaggtggggctcctcttgtcatcagctcccagaggagcacgccaaaggaccacacatctgacttggtggtaaacttttgagtttgcagactttccaaagccatccacttcactggcagctttgcacctgttttgttgtgtacactatagtattctttatcatacatgtctctggcaagaccaaaatcagcaaccttgactgtgaatttttcatccagcatacagtttcttgcagccaagtctctgtggacaaactttttgcttgcaagatatttcatgcctttggctacttgaagaccaaagccaataagatcttttacagttggattatgagtctcatttcgaatgaaatttcgaagatctccatgtttcatgtatggtaggaccaccagcggagacccttcacttcgcaggcagattcccaggagcgagaggacattgggatgactaaaatctttcatgatgattccctcggtcagaaattgggaaacttctcctatgtcagtgattctgttcaaggatttcacagcacagtgaattttcttgccatcattgtccaacaaagtcccatgatatacacaaccaaaatgccctcttcctatgacttcattgaaatgcacaatcaggctactgggcccaatcactacatgctgcactgcctggaccagctctggatttagagcactgaggtcaatgtggacagtattttgcagtaatggactggatatatcagagtccccactagttaggatgggggacatgtctgtcagaggatactgcacttgtcggcatgaaccgttctgagatgaattaggaaactgatcttctggaaaagtagctcggtagtctacagattcatttgaaaccatttctgtagttgggcttacacttcgggcacttacaagcctatccaaatgaggagtgtgtactcttgcatcgtagcgaactaattcactgcccagatctttaatttgctttctctttttcagccacaggaaaaacccaagtagtaataacagtgctgttgatattgagacaacaccagcaatcaatcctgtgaaattctgatctggttgaactattacttttccaaggacggttgaagaaattgcttgcttccactctatatttagctcgctgttcaatttcagcaggtcattggggaccgtgcataaaacggcttcagaatgtaagtgtatattctcacagctcttatttccaacttttaacacttcacctttaactgcttcagggtcaatatcatttcccttaatttccagtacattttcattgcccattgagatcatcactggcttttcaaaaggcttaaacacaggattatgtacataaatgagatcaaagtatttggaaaggatcccatctaacatgaaaaaggctttggttttcagggggagttgcagattcagctgttgcagggaaggagtggtacaacagattatctctgaattagagcgatgttgacatgcctaataaaagatttggttggatgaatttcatagacaatgggatcttcacggtaactgaagatgcttgtctctcggttggctaagtcaattttcaatttaacagcaaactcagttgaaatggtttgggctggggtataacattcaagaatactgtttgacacactttttaaagtacatgtttttccaccaattgaaatgtgtctagaattcccactgtttaggtaatttccagttaaagtaagtaaagtgccaccagccataggaccgtatttcggcgaaatacttgttattacaggatccacataggagaatgtactgtattgtgttgtcccgtggccatttgaaataattatggacatattgaaatgcttattcatggcaggaccaactgtgcatttcaatgtattcatcgtgctctcacttaaagtcaaggtgcagctctcatttccaaggagaactctagttttctttaaatcaaatttattattcctccgaaatccaaagtcccagccacatatggtcagccttgtccctccttcaaggggtgcactatttgggaaaaccttgtagattgcaggcagacagatctgttgagtccatgtcccgctcaggcattcctccgatcgcacacatttgtcgtggcaccagccacactgaacaaagggtggggcagagaggcattgactgcaggactggaaatgtctgcagcccaagccattcaatgggatcttcgtgatcttcttcccagtgataaccagtgtgtagccattttggtttaatgtatgctccacaatcacttctggagacactggatgggagtccaggagaaaattcacatgaggggttgatggtcctgatcgagaaaccacaacctgcatgaagcgaccctctgatgtcccaagattagctatggtgaggtctcctttaatgaaggtggatatagatgttaagaggacttcgctgaattgacccatgaataagtcaacgcgctgcaaagctgtggtaaactctgttcgatattcatcacggcgcgcttcacagcctgatgaatttctcagaagtgtcctattaaagcagtgctcatgattgggtccgtaaaaatgctggagacatctcacattgtttttgttgacgatcttgttgaagaagtcgttgacatatttgatagggaatgcacacatggcagatcgatccattggttcggcagaatctggcttgctttgtgcgaacaccccgaaaagaatgtcatcattcaggctggctcctatttgtctagcaagctgggccccaggcttgctgacatacgcagcctgaagtatattaaacacttccttctttgtggatctcttttttctcttttctgtgagaatacactccagaggcatttccatgtaggaatgcaatccagagtttatggaacagaacctgattattcttgtgtgaaaagtctgagcatctagagtttccctttggaccgtcaagaagtaaataaaattgttgctttcaaaggcatggacatacttaatggggtaagaatctctgaactcaggtaaaacatcaatgtaggactggtccgtcaaaaacataaaaccatctttcgtttcctttagccttctcactgatatcgaatgcaatggatgatctgggaaataagaagaatttatggtattgcctacaaagaagttgatgaaccggtcctttacagatgaaaggactttggctcccagggcgctcaccacacagtcaggacactggctgggctcttctatctgtggggagaatatgcagtgaacctccgactgtatgtcagcagtatgattgtggggaaagacatgtcgctggcaggtccctctgttgacgctgccacagctaatgagttgatcatcatagtaggtgtcgacaactagagccatgttgatgttatctttccaaacacctcctgataaattggctttgctgctgcagtcctgacatgggaaacaatctgggtgttccagcacaggcccagtcttgtactcagcaaccttctgaaggtcttcctcatttaaaacataaatgtagttagtggcaccaaggaaaatgtgatgctcatgtagaatgacattctggatgggtgtttccgcggtgaagttgggaagctgatacttcatattcacattcatctcggactttgctagtgcctctttacactccccattgctcctctgcaccaaggtaaacaggagcacgaggatgccaggtgcaagcacagcgggggccttcattatgagaggtttatctttcggtgcccaggaaccagtggagaagtcagcggcgcaaggaccacacgcgcgctccgcgcctccccgcctcctctcagcaagtcagctgtcgccccgcatctggctcgcgccctccactcggctccgcatctgctcacaaagcgctcggggcgccgcgggcggcgagggcctccgggtcacctgc。
其中，所述第一引物对只要能特异性地扩增SEQ ID NO.1的cDNA片段即可，优选地，所述第一引物对可以含有如SEQ ID NO:4所示的第一引物和SEQ ID NO:5所示的第二引物。第一探针只要能与SEQ ID NO.1的cDNA片段特异性杂交即可，优选地，所述第一探针的序列中可以含有如SEQ ID NO:6所示的序列。
优选地，SEQ ID NO:4所示的序列可以为：
atccaaccaaatcttttattaggcatg；
SEQ ID NO:5所示的序列可以为：
cttctggaaaagtagctcggtagtct；
SEQ ID NO:6所示的序列可以为：
caaatcttttattaggcatgtcaacatc。
本公开的第二方面提供一种评价胶质瘤和/或胃腺癌预后性的试剂盒，该试剂盒包括能够特异性地扩增SEQ ID NO.2的mRNA片段的第二引物对，和/或，能够与SEQ ID NO.2的mRNA片段特异性杂交的第二探针，其中，所述SEQ ID NO:2的mRNA片段至少含有SEQ IDNO:2的第2452-2469位、第2464-2481位或者第2458-2474位的mRNA序列。
优选地，所述SEQ ID NO:2的mRNA片段至少含有SEQ ID NO:2第2416-2516位的mRNA序列；优选地，所述SEQ ID NO:2的mRNA片段是指如SEQ ID NO:2所示的mRNA序列。
其中，SEQ ID NO:2所示的mRNA序列为与SEQ ID NO:1所示的cDNA序列互补的RNA序列，SEQ ID NO:2所示的序列如下所示：
gcaggugacccggaggcccucgccgcccgcggcgccccgagcgcuuugugagcagaugcggagccgaguggagggcgcgagccagaugcggggcgacagcugacuugcugagaggaggcggggaggcgcggagcgcgcgugugguccuugcgccgcugacuucuccacugguuccugggcaccgaaagauaaaccucucauaaugaaggcccccgcugugcuugcaccuggcauccucgugcuccuguuuaccuuggugcagaggagcaauggggaguguaaagaggcacuagcaaaguccgagaugaaugugaauaugaaguaucagcuucccaacuucaccgcggaaacacccauccagaaugucauucuacaugagcaucacauuuuccuuggugccacuaacuacauuuauguuuuaaaugaggaagaccuucagaagguugcugaguacaagacugggccugugcuggaacacccagauuguuucccaugucaggacugcagcagcaaagccaauuuaucaggagguguuuggaaagauaacaucaacauggcucuaguugucgacaccuacuaugaugaucaacucauuagcuguggcagcgucaacagagggaccugccagcgacaugucuuuccccacaaucauacugcugacauacagucggagguucacugcauauucuccccacagauagaagagcccagccaguguccugacuguguggugagcgcccugggagccaaaguccuuucaucuguaaaggaccgguucaucaacuucuuuguaggcaauaccauaaauucuucuuauuucccagaucauccauugcauucgauaucagugagaaggcuaaaggaaacgaaagaugguuuuauguuuuugacggaccaguccuacauugauguuuuaccugaguucagagauucuuaccccauuaaguauguccaugccuuugaaagcaacaauuuuauuuacuucuugacgguccaaagggaaacucuagaugcucagacuuuucacacaagaauaaucagguucuguuccauaaacucuggauugcauuccuacauggaaaugccucuggaguguauucucacagaaaagagaaaaaagagauccacaaagaaggaaguguuuaauauacuucaggcugcguaugucagcaagccuggggcccagcuugcuagacaaauaggagccagccugaaugaugacauucuuuucgggguguucgcacaaagcaagccagauucugccgaaccaauggaucgaucugccaugugugcauucccuaucaaauaugucaacgacuucuucaacaagaucgucaacaaaaacaaugugagaugucuccagcauuuuuacggacccaaucaugagcacugcuuuaauaggacacuucugagaaauucaucaggcugugaagcgcgccgugaugaauaucgaacagaguuuaccacagcuuugcagcgcguugacuuauucaugggucaauucagcgaaguccucuuaacaucuauauccaccuucauuaaaggagaccucaccauagcuaaucuugggacaucagagggucgcuucaugcagguugugguuucucgaucaggaccaucaaccccucaugugaauuuucuccuggacucccauccagugucuccagaagugauuguggagcauacauuaaaccaaaauggcuacacacugguuaucacugggaagaagaucacgaagaucccauugaauggcuugggcugcagacauuuccaguccugcagucaaugccucucugccccacccuuuguucaguguggcuggugccacgacaaaugugugcgaucggaggaaugccugagcgggacauggacucaacagaucugucugccugcaaucuacaagguuuucccaaauagugcaccccuugaaggagggacaaggcugaccauauguggcugggacuuuggauuucggaggaauaauaaauuugauuuaaagaaaacuagaguucuccuuggaaaugagagcugcaccuugacuuuaagugagagcacgaugaauacauugaaaugcacaguugguccugccaugaauaagcauuucaauauguccauaauuauuucaaauggccacgggacaacacaauacaguacauucuccuauguggauccuguaauaacaaguauuucgccgaaauacgguccuauggcugguggcacuuuacuuacuuuaacuggaaauuaccuaaacagugggaauucuagacacauuucaauugguggaaaaacauguacuuuaaaaagugugucaaacaguauucuugaauguuauaccccagcccaaaccauuucaacugaguuugcuguuaaauugaaaauugacuuagccaaccgagagacaagcaucuucaguuaccgugaagaucccauugucuaugaaauucauccaaccaaaucuuuuauuaggcaugucaacaucgcucuaauucagagauaaucuguuguaccacuccuucccugcaacagcugaaucugcaacucccccugaaaaccaaagccuuuuucauguuagaugggauccuuuccaaauacuuugaucucauuuauguacauaauccuguguuuaagccuuuugaaaagccagugaugaucucaaugggcaaugaaaauguacuggaaauuaagggaaaugauauugacccugaagcaguuaaaggugaaguguuaaaaguuggaaauaagagcugugagaauauacacuuacauucugaagccguuuuaugcacgguccccaaugaccugcugaaauugaacagcgagcuaaauauagaguggaagcaagcaauuucuucaaccguccuuggaaaaguaauaguucaaccagaucagaauuucacaggauugauugcugguguugucucaauaucaacagcacuguuauuacuacuuggguuuuuccuguggcugaaaaagagaaagcaaauuaaagaucugggcagugaauuaguucgcuacgaugcaagaguacacacuccucauuuggauaggcuuguaagugcccgaaguguaagcccaacuacagaaaugguuucaaaugaaucuguagacuaccgagcuacuuuuccagaagaucaguuuccuaauucaucucagaacgguucaugccgacaagugcaguauccucugacagacaugucccccauccuaacuaguggggacucugauauauccaguccauuacugcaaaauacuguccacauugaccucagugcucuaaauccagagcugguccaggcagugcagcauguagugauugggcccaguagccugauugugcauuucaaugaagucauaggaagagggcauuuugguuguguauaucaugggacuuuguuggacaaugauggcaagaaaauucacugugcugugaaauccuugaacagaaucacugacauaggagaaguuucccaauuucugaccgagggaaucaucaugaaagauuuuagucaucccaauguccucucgcuccugggaaucugccugcgaagugaagggucuccgcuggugguccuaccauacaugaaacauggagaucuucgaaauuucauucgaaaugagacucauaauccaacuguaaaagaucuuauuggcuuuggucuucaaguagccaaaggcaugaaauaucuugcaagcaaaaaguuuguccacagagacuuggcugcaagaaacuguaugcuggaugaaaaauucacagucaagguugcugauuuuggucuugccagagacauguaugauaaagaauacuauaguguacacaacaaaacaggugcaaagcugccagugaaguggauggcuuuggaaagucugcaaacucaaaaguuuaccaccaagucagaugugugguccuuuggcgugcuccucugggagcugaugacaagaggagccccaccuuauccugacguaaacaccuuugauauaacuguuuacuuguugcaagggagaagacuccuacaacccgaauacugcccagaccccuuauaugaaguaaugcuaaaaugcuggcacccuaaagccgaaaugcgcccauccuuuucugaacuggugucccggauaucagcgaucuucucuacuuucauuggggagcacuauguccaugugaacgcuacuuaugugaacguaaaaugugucgcuccguauccuucucuguugucaucagaagauaacgcugaugaugagguggacacacgaccagccuccuucugggagacaucauagugcuaguacuaugucaaagcaacaguccacacuuuguccaaugguuuuuucacugccugaccuuuaaaaggccaucgauauucuuugcucuugccaaaauugcacuauuauaggacuuguauuguuauuuaaauuacuggauucuaaggaauuucuuaucugacagagcaucagaaccagaggcuuggucccacaggccacggaccaauggccugcagccgugacaacacuccugucauauuggaguccaaaacuugaauucuggguugaauuuuuuaaaaaucagguaccacuugauuucauaugggaaauugaagcaggaaauauugagggcuucuugaucacagaaaacucagaagagauaguaaugcucaggacaggagcggcagccccagaacaggccacucauuuagaauucuaguguuucaaaacacuuuuguguguuguauggucaauaacauuuuucauuacugauggugucauucacccauuagguaaacauucccuuuuaaauguuuguuuguuuuuugagacaggaucucacucuguugccagggcuguagugcaguggugugaucauagcucacugcaaccuccaccucccaggcucaagccucccgaauagcugggacuacaggcgcacaccaccauccccggcuaauuuuuguauuuuuuguagagacgggguuuugccauguugccaaggcugguuucaaacuccuggacucaagaaauccacccaccucagccucccaaagugcuaggauuacaggcaugagccacugcgcccagcccuuauaaauuuuuguauagacauuccuuugguuggaagaauauuuauaggcaauacagucaaaguuucaaaauagcaucacacaaaacauguuuauaaaugaacaggauguaauguacauagaugacauuaagaaaauuuguaugaaauaauuuagucaucaugaaauauuuaguugucauauaaaaacccacuguuugagaaugaugcuacucugaucuaaugaaugugaacauguagauguuuuguguguauuuuuuuaaaugaaaacucaaaauaagacaaguaauuuguugauaaauauuuuuaaagauaacucagcauguuuguaaagcaggauacauuuuacuaaaagguucauugguuccaaucacagcucauagguagagcaaagaaaggguggauggauugaaaagauuagccucugucucgguggcagguucccaccucgcaagcaauuggaaacaaaacuuuuggggaguuuuauuuugcauuaggguguguuuuauguuaagcaaaacauacuuuagaaacaaaugaaaaaggcaauugaaaaucccagcuauuucaccuagauggaauagccacccugagcagaacuuugugaugcuucauucuguggaauuuugugcuugcuacuguauagugcaugugguguagguuacucuaacugguuuugucgacguaaacauuuaaaguguuauauuuuuuauaaaaauguuuauuuuuaaugauaugagaaaaauuuuguuaggccacaaaaacacugcacugugaacauuuuagaaaagguaugucagacugggauuaaugacagcaugauuuucaaugacuguaaauugcgauaaggaaauguacugauugccaauacaccccacccucauuacaucaucaggacuugaagccaaggguuaacccagcaagcuacaaagagggugugucacacugaaacucaauaguugaguuuggcuguuguugcaggaaaaugauuauaacuaaaagcucucugauagugcagagacuuaccagaagacacaaggaauuguacugaagagcuauuacaauccaaauauugccguuucauaaauguaauaaguaauacuaauucacagaguauuguaaaugguggaugacaaaagaaaaucugcucuguggaaagaaagaacugucucuaccagggucaagagcaugaacgcaucaauagaaagaacucggggaaacaucccaucaacaggacuacacacuuguauauacauucuugagaacacugcaaugugaaaaucacguuugcuauuuauaaacuuguccuuagauuaaugugucuggacagauugugggaguaagugauucuucuaagaauuagauacuugucacugccuauaccugcagcugaacugaaugguacuucguauguuaauaguuguucugauaaaucaugcaauuaaaguaaagugaugcaacaucuugua。
其中，所述第二引物对只要能特异性地扩增SEQ ID NO.2的mRNA片段即可，优选地，所述第二引物对可以含有SEQ ID NO:7所示的第三引物和如SEQ ID NO:8所示的第四引物。第二探针只要能与SEQ ID NO.2的mRNA片段特异性杂交即可，优选地，所述第二探针的序列中可以含有如SEQ ID NO:9所示的序列。
优选地，SEQ ID NO:7所示的序列可以为：
taatacgactcactatagggagcgatgttgacatgcctaataaaaga；
SEQ ID NO:8所示的序列可以为：
gtgtgtcaaacagtattcttg；
SEQ ID NO:9所示的序列可以为：
ttgacatgcctaataaaagatttggttg。
本公开的第三方面提供一种评价胶质瘤和/或胃腺癌预后性的试剂盒，该试剂盒包括抗SEQ ID NO.3的氨基酸片段的抗体，其中，所述SEQ ID NO.3的氨基酸片段至少含有SEQ ID NO:3的第750-764位的氨基酸序列。
优选地，所述SEQ ID NO.3的氨基酸片段至少含有SEQ ID NO:3的第722-764位的氨基酸序列；更优选地，所述SEQ ID NO.3的氨基酸片段是指如SEQ ID NO:3所示的氨基酸序列。
其中，SEQ ID NO:3所示的氨基酸序列为由SEQ ID NO:2所示的mRNA序列编码的氨基酸序列，SEQ ID NO:3所示的氨基酸序列如下所示：
MKAPAVLAPGILVLLFTLVQRSNGECKEALAKSEMNVNMKYQLPNFTAETPIQNVILHEHHIFLGATNYIYVLNEEDLQKVAEYKTGPVLEHPDCFPCQDCSSKANLSGGVWKDNINMALVVDTYYDDQLISCGSVNRGTCQRHVFPHNHTADIQSEVHCIFSPQIEEPSQCPDCVVSALGAKVLSSVKDRFINFFVGNTINSSYFPDHPLHSISVRRLKETKDGFMFLTDQSYIDVLPEFRDSYPIKYVHAFESNNFIYFLTVQRETLDAQTFHTRIIRFCSINSGLHSYMEMPLECILTEKRKKRSTKKEVFNILQAAYVSKPGAQLARQIGASLNDDILFGVFAQSKPDSAEPMDRSAMCAFPIKYVNDFFNKIVNKNNVRCLQHFYGPNHEHCFNRTLLRNSSGCEARRDEYRTEFTTALQRVDLFMGQFSEVLLTSISTFIKGDLTIANLGTSEGRFMQVVVSRSGPSTPHVNFLLDSHPVSPEVIVEHTLNQNGYTLVITGKKITKIPLNGLGCRHFQSCSQCLSAPPFVQCGWCHDKCVRSEECLSGTWTQQICLPAIYKVFPNSAPLEGGTRLTICGWDFGFRRNNKFDLKKTRVLLGNESCTLTLSESTMNTLKCTVGPAMNKHFNMSIIISNGHGTTQYSTFSYVDPVITSISPKYGPMAGGTLLTLTGNYLNSGNSRHISIGGKTCTLKSVSNSILECYTPAQTISTEFAVKLKIDLANRETSIFSYREDPIVYEIHPTKSFIRHVNIALIQR。
其中，抗SEQ ID NO.3的氨基酸片段的抗体可以为单克隆抗体和/或多克隆抗体。
本公开的第四方面提供一种分子试剂在制备评价胶质瘤和/或胃腺癌预后性的试剂盒中的用途，所述分子试剂包括如下(1)-(8)中的至少一种：
(1)如SEQ ID NO.1所示的cDNA；
(2)能够特异性地扩增如SEQ ID NO.1所示的cDNA的第一引物对；
(3)能够与如SEQ ID NO.1所示的cDNA特异性杂交的第一探针；
(4)如SEQ ID NO.2所示的mRNA；
(5)能够特异性地扩增如SEQ ID NO.2所示的mRNA的第二引物对；
(6)能够与如SEQ ID NO.2所示的mRNA特异性杂交的第二探针；
(7)如SEQ ID NO.3所示的蛋白；
(8)抗如SEQ ID NO.3所示的蛋白的抗体。
本公开的第五方面提供一种评价胶质瘤和/或胃腺癌预后性的系统，该系统包括扩增装置、测序装置、计算装置和输出装置；所述扩增装置含有采集单元和扩增单元，所述采集单元用于采集模板核酸片段和扩增引物，所述扩增单元用于利用所述扩增引物对所述模板核酸片段进行扩增，得到扩增产物；所述测序装置用于对所述扩增产物进行核酸序列测序，得到扩增产物序列；所述计算装置包括存储器和处理器，所述存储器中存储有计算机程序，所述处理器被配置为执行所述存储器中存储的计算机程序，以实现如下判别：若所述扩增产物序列中含有如SEQ ID NO.1所示的cDNA的序列和/或如SEQ ID NO.2所示的mRNA的序列，则判定所述模板核酸片段对应的胶质瘤和/或胃腺癌预后性较差；所述输出装置用于输出所述计算装置的判定结果。
本公开的第六方面提供一种评价胶质瘤和/或胃腺癌预后性的方法，该方法包括检测待测胶质瘤和/或胃腺癌样本中是否含有如上所述的SEQ ID NO.1的cDNA片段和/或SEQ ID NO.2的mRNA片段和/或SEQ ID NO.3的氨基酸片段的步骤，如果待测胶质瘤和/或胃腺癌样本中含有如上所述的SEQ ID NO.1的cDNA片段和/或SEQ ID NO.2的mRNA片段和/或SEQ ID NO.3的氨基酸片段，则指示该待测胶质瘤和/或胃腺癌样本对应的胶质瘤和/或胃腺癌患者的预后性更差。其中，可以通过PCR、RT-RPA、核酸杂交或者高通量测序的方法检测上述cDNA和/或mRNA，可以通过免疫杂交检测上述蛋白。
以下通过实施例进一步详细说明本公开。
制备实施例
本制备实施例用于获取胶质瘤样本，并获取其中的总RNA和总cDNA。
使用符合医学伦理委员会标准的操作，收集胶质瘤的样本1211例，并判断每一例胶质瘤样本的病理学特征。其中，收集样本的每一位病人都在收集样本之前得到本人及其治疗专家的同意，并具有书面的证明材料。其中，利用病理学诊断方法诊断胶质瘤，并根据胶质瘤样本对应患者的总生存期来评估胶质瘤样本的预后性，患者总生存期越长，则胶质瘤的预后性越好。胶质瘤样本对应患者的性别、年龄、病理学分级、肿瘤类别、以及总生存期等特征见表1。
表1
使用DNA提取试剂盒(购自Qiagen)，按照其使用说明书提取上述胶质瘤样本的总RNA。该总RNA经完整性分析仪检测，确认其RNA完整性指数(RNA Integrity Number,RIN)大于7.0。使用反转录试剂盒(购自Invitrogen)按照其使用说明书以该总RNA为模板合成双链cDNA。
实施例1
本实施例对制备实施例合成得到的胶质瘤样本的cDNA进行PCR验证。
PCR验证所用的引物为如SEQ ID NO:4所示的第一引物和如SEQ ID NO:5所示的第二引物。PCR的操作按照合成引物和PCR试剂盒的说明书进行。PCR的产物经过琼脂糖凝胶核酸电泳展示扩增条带的有无，如图1所示，所出现的扩增条带使用DNA凝胶回收试剂盒(QIAquick PCR purification kit，购自Qiagen)进行回收，然后进行Sanger测序，测序结果见图2。
由图1和图2可知，胶质瘤样品中存在10号外显子缺失的MET基因。
实施例2
本实施例对制备实施例收集得到的胶质瘤样本的RNA进行测序。
将各个样本的RNA使用RNA文库构建试剂盒(购自Illumina)构建RNA文库，然后使用测序平台(IlluminaHiSeq 2000)对RNA文库进行RNA测序，并筛选出RNA测序结果中含有如SEQ ID NO:2所示的mRNA的胶质瘤样本，结果见表2。
表2
由表2可以看出，检测到含有如SEQ ID NO:2所示的mRNA的胶质瘤样本有14例，其中位生存期为200天，未检测到如SEQ ID NO:2所示的mRNA的胶质瘤样本有1197例，其中位生存期为872天，说明含有SEQ ID NO:2所示的mRNA的胶质瘤预后较差，由此可以用于评价胶质瘤的预后性。
此外，分别在全级别胶质瘤、IDH突变的全级别胶质瘤、继发胶质母细胞瘤和IDH突变的胶质母细胞瘤中，通过生存曲线(Kaplan–Meiercurve)对含有如SEQ ID NO:2所示的mRNA的胶质瘤样本和不含有如SEQ ID NO:2所示的mRNA的胶质瘤样本的整体生存情况进行了比较，结果均显示含有如SEQ ID NO:2所示的mRNA的胶质瘤样本整体生存较差，见图3-6。
同理可以证明，含有如SEQ ID NO:2所示的mRNA的胃腺癌样本对应患者的整体生存较差，见图7。
以上结合附图详细描述了本公开的优选实施方式，但是，本公开并不限于上述实施方式中的具体细节，在本公开的技术构思范围内，可以对本公开的技术方案进行多种简单变型，这些简单变型均属于本公开的保护范围。
另外需要说明的是，在上述具体实施方式中所描述的各个具体技术特征，在不矛盾的情况下，可以通过任何合适的方式进行组合。
此外，本公开的各种不同的实施方式之间也可以进行任意组合，只要其不违背本公开的思想，其同样应当视为本公开所公开的内容。
序列表
<110> 北京市神经外科研究所
<120> 评价胶质瘤和/或胃腺癌预后性的试剂盒和系统
<130> 16125BJNI-CRC
<160> 9
<170> SIPOSequenceListing 1.0
<210> 1
<211> 6537
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 1
tacaagatgt tgcatcactt tactttaatt gcatgattta tcagaacaac tattaacata 60
cgaagtacca ttcagttcag ctgcaggtat aggcagtgac aagtatctaa ttcttagaag 120
aatcacttac tcccacaatc tgtccagaca cattaatcta aggacaagtt tataaatagc 180
aaacgtgatt ttcacattgc agtgttctca agaatgtata tacaagtgtg tagtcctgtt 240
gatgggatgt ttccccgagt tctttctatt gatgcgttca tgctcttgac cctggtagag 300
acagttcttt ctttccacag agcagatttt cttttgtcat ccaccattta caatactctg 360
tgaattagta ttacttatta catttatgaa acggcaatat ttggattgta atagctcttc 420
agtacaattc cttgtgtctt ctggtaagtc tctgcactat cagagagctt ttagttataa 480
tcattttcct gcaacaacag ccaaactcaa ctattgagtt tcagtgtgac acaccctctt 540
tgtagcttgc tgggttaacc cttggcttca agtcctgatg atgtaatgag ggtggggtgt 600
attggcaatc agtacatttc cttatcgcaa tttacagtca ttgaaaatca tgctgtcatt 660
aatcccagtc tgacatacct tttctaaaat gttcacagtg cagtgttttt gtggcctaac 720
aaaatttttc tcatatcatt aaaaataaac atttttataa aaaatataac actttaaatg 780
tttacgtcga caaaaccagt tagagtaacc tacaccacat gcactataca gtagcaagca 840
caaaattcca cagaatgaag catcacaaag ttctgctcag ggtggctatt ccatctaggt 900
gaaatagctg ggattttcaa ttgccttttt catttgtttc taaagtatgt tttgcttaac 960
ataaaacaca ccctaatgca aaataaaact ccccaaaagt tttgtttcca attgcttgcg 1020
aggtgggaac ctgccaccga gacagaggct aatcttttca atccatccac cctttctttg 1080
ctctacctat gagctgtgat tggaaccaat gaacctttta gtaaaatgta tcctgcttta 1140
caaacatgct gagttatctt taaaaatatt tatcaacaaa ttacttgtct tattttgagt 1200
tttcatttaa aaaaatacac acaaaacatc tacatgttca cattcattag atcagagtag 1260
catcattctc aaacagtggg tttttatatg acaactaaat atttcatgat gactaaatta 1320
tttcatacaa attttcttaa tgtcatctat gtacattaca tcctgttcat ttataaacat 1380
gttttgtgtg atgctatttt gaaactttga ctgtattgcc tataaatatt cttccaacca 1440
aaggaatgtc tatacaaaaa tttataaggg ctgggcgcag tggctcatgc ctgtaatcct 1500
agcactttgg gaggctgagg tgggtggatt tcttgagtcc aggagtttga aaccagcctt 1560
ggcaacatgg caaaaccccg tctctacaaa aaatacaaaa attagccggg gatggtggtg 1620
tgcgcctgta gtcccagcta ttcgggaggc ttgagcctgg gaggtggagg ttgcagtgag 1680
ctatgatcac accactgcac tacagccctg gcaacagagt gagatcctgt ctcaaaaaac 1740
aaacaaacat ttaaaaggga atgtttacct aatgggtgaa tgacaccatc agtaatgaaa 1800
aatgttattg accatacaac acacaaaagt gttttgaaac actagaattc taaatgagtg 1860
gcctgttctg gggctgccgc tcctgtcctg agcattacta tctcttctga gttttctgtg 1920
atcaagaagc cctcaatatt tcctgcttca atttcccata tgaaatcaag tggtacctga 1980
tttttaaaaa attcaaccca gaattcaagt tttggactcc aatatgacag gagtgttgtc 2040
acggctgcag gccattggtc cgtggcctgt gggaccaagc ctctggttct gatgctctgt 2100
cagataagaa attccttaga atccagtaat ttaaataaca atacaagtcc tataatagtg 2160
caattttggc aagagcaaag aatatcgatg gccttttaaa ggtcaggcag tgaaaaaacc 2220
attggacaaa gtgtggactg ttgctttgac atagtactag cactatgatg tctcccagaa 2280
ggaggctggt cgtgtgtcca cctcatcatc agcgttatct tctgatgaca acagagaagg 2340
atacggagcg acacatttta cgttcacata agtagcgttc acatggacat agtgctcccc 2400
aatgaaagta gagaagatcg ctgatatccg ggacaccagt tcagaaaagg atgggcgcat 2460
ttcggcttta gggtgccagc attttagcat tacttcatat aaggggtctg ggcagtattc 2520
gggttgtagg agtcttctcc cttgcaacaa gtaaacagtt atatcaaagg tgtttacgtc 2580
aggataaggt ggggctcctc ttgtcatcag ctcccagagg agcacgccaa aggaccacac 2640
atctgacttg gtggtaaact tttgagtttg cagactttcc aaagccatcc acttcactgg 2700
cagctttgca cctgttttgt tgtgtacact atagtattct ttatcataca tgtctctggc 2760
aagaccaaaa tcagcaacct tgactgtgaa tttttcatcc agcatacagt ttcttgcagc 2820
caagtctctg tggacaaact ttttgcttgc aagatatttc atgcctttgg ctacttgaag 2880
accaaagcca ataagatctt ttacagttgg attatgagtc tcatttcgaa tgaaatttcg 2940
aagatctcca tgtttcatgt atggtaggac caccagcgga gacccttcac ttcgcaggca 3000
gattcccagg agcgagagga cattgggatg actaaaatct ttcatgatga ttccctcggt 3060
cagaaattgg gaaacttctc ctatgtcagt gattctgttc aaggatttca cagcacagtg 3120
aattttcttg ccatcattgt ccaacaaagt cccatgatat acacaaccaa aatgccctct 3180
tcctatgact tcattgaaat gcacaatcag gctactgggc ccaatcacta catgctgcac 3240
tgcctggacc agctctggat ttagagcact gaggtcaatg tggacagtat tttgcagtaa 3300
tggactggat atatcagagt ccccactagt taggatgggg gacatgtctg tcagaggata 3360
ctgcacttgt cggcatgaac cgttctgaga tgaattagga aactgatctt ctggaaaagt 3420
agctcggtag tctacagatt catttgaaac catttctgta gttgggctta cacttcgggc 3480
acttacaagc ctatccaaat gaggagtgtg tactcttgca tcgtagcgaa ctaattcact 3540
gcccagatct ttaatttgct ttctcttttt cagccacagg aaaaacccaa gtagtaataa 3600
cagtgctgtt gatattgaga caacaccagc aatcaatcct gtgaaattct gatctggttg 3660
aactattact tttccaagga cggttgaaga aattgcttgc ttccactcta tatttagctc 3720
gctgttcaat ttcagcaggt cattggggac cgtgcataaa acggcttcag aatgtaagtg 3780
tatattctca cagctcttat ttccaacttt taacacttca cctttaactg cttcagggtc 3840
aatatcattt cccttaattt ccagtacatt ttcattgccc attgagatca tcactggctt 3900
ttcaaaaggc ttaaacacag gattatgtac ataaatgaga tcaaagtatt tggaaaggat 3960
cccatctaac atgaaaaagg ctttggtttt cagggggagt tgcagattca gctgttgcag 4020
ggaaggagtg gtacaacaga ttatctctga attagagcga tgttgacatg cctaataaaa 4080
gatttggttg gatgaatttc atagacaatg ggatcttcac ggtaactgaa gatgcttgtc 4140
tctcggttgg ctaagtcaat tttcaattta acagcaaact cagttgaaat ggtttgggct 4200
ggggtataac attcaagaat actgtttgac acacttttta aagtacatgt ttttccacca 4260
attgaaatgt gtctagaatt cccactgttt aggtaatttc cagttaaagt aagtaaagtg 4320
ccaccagcca taggaccgta tttcggcgaa atacttgtta ttacaggatc cacataggag 4380
aatgtactgt attgtgttgt cccgtggcca tttgaaataa ttatggacat attgaaatgc 4440
ttattcatgg caggaccaac tgtgcatttc aatgtattca tcgtgctctc acttaaagtc 4500
aaggtgcagc tctcatttcc aaggagaact ctagttttct ttaaatcaaa tttattattc 4560
ctccgaaatc caaagtccca gccacatatg gtcagccttg tccctccttc aaggggtgca 4620
ctatttggga aaaccttgta gattgcaggc agacagatct gttgagtcca tgtcccgctc 4680
aggcattcct ccgatcgcac acatttgtcg tggcaccagc cacactgaac aaagggtggg 4740
gcagagaggc attgactgca ggactggaaa tgtctgcagc ccaagccatt caatgggatc 4800
ttcgtgatct tcttcccagt gataaccagt gtgtagccat tttggtttaa tgtatgctcc 4860
acaatcactt ctggagacac tggatgggag tccaggagaa aattcacatg aggggttgat 4920
ggtcctgatc gagaaaccac aacctgcatg aagcgaccct ctgatgtccc aagattagct 4980
atggtgaggt ctcctttaat gaaggtggat atagatgtta agaggacttc gctgaattga 5040
cccatgaata agtcaacgcg ctgcaaagct gtggtaaact ctgttcgata ttcatcacgg 5100
cgcgcttcac agcctgatga atttctcaga agtgtcctat taaagcagtg ctcatgattg 5160
ggtccgtaaa aatgctggag acatctcaca ttgtttttgt tgacgatctt gttgaagaag 5220
tcgttgacat atttgatagg gaatgcacac atggcagatc gatccattgg ttcggcagaa 5280
tctggcttgc tttgtgcgaa caccccgaaa agaatgtcat cattcaggct ggctcctatt 5340
tgtctagcaa gctgggcccc aggcttgctg acatacgcag cctgaagtat attaaacact 5400
tccttctttg tggatctctt ttttctcttt tctgtgagaa tacactccag aggcatttcc 5460
atgtaggaat gcaatccaga gtttatggaa cagaacctga ttattcttgt gtgaaaagtc 5520
tgagcatcta gagtttccct ttggaccgtc aagaagtaaa taaaattgtt gctttcaaag 5580
gcatggacat acttaatggg gtaagaatct ctgaactcag gtaaaacatc aatgtaggac 5640
tggtccgtca aaaacataaa accatctttc gtttccttta gccttctcac tgatatcgaa 5700
tgcaatggat gatctgggaa ataagaagaa tttatggtat tgcctacaaa gaagttgatg 5760
aaccggtcct ttacagatga aaggactttg gctcccaggg cgctcaccac acagtcagga 5820
cactggctgg gctcttctat ctgtggggag aatatgcagt gaacctccga ctgtatgtca 5880
gcagtatgat tgtggggaaa gacatgtcgc tggcaggtcc ctctgttgac gctgccacag 5940
ctaatgagtt gatcatcata gtaggtgtcg acaactagag ccatgttgat gttatctttc 6000
caaacacctc ctgataaatt ggctttgctg ctgcagtcct gacatgggaa acaatctggg 6060
tgttccagca caggcccagt cttgtactca gcaaccttct gaaggtcttc ctcatttaaa 6120
acataaatgt agttagtggc accaaggaaa atgtgatgct catgtagaat gacattctgg 6180
atgggtgttt ccgcggtgaa gttgggaagc tgatacttca tattcacatt catctcggac 6240
tttgctagtg cctctttaca ctccccattg ctcctctgca ccaaggtaaa caggagcacg 6300
aggatgccag gtgcaagcac agcgggggcc ttcattatga gaggtttatc tttcggtgcc 6360
caggaaccag tggagaagtc agcggcgcaa ggaccacacg cgcgctccgc gcctccccgc 6420
ctcctctcag caagtcagct gtcgccccgc atctggctcg cgccctccac tcggctccgc 6480
atctgctcac aaagcgctcg gggcgccgcg ggcggcgagg gcctccgggt cacctgc 6537
<210> 2
<211> 6537
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 2
gcaggugacc cggaggcccu cgccgcccgc ggcgccccga gcgcuuugug agcagaugcg 60
gagccgagug gagggcgcga gccagaugcg gggcgacagc ugacuugcug agaggaggcg 120
gggaggcgcg gagcgcgcgu gugguccuug cgccgcugac uucuccacug guuccugggc 180
accgaaagau aaaccucuca uaaugaaggc ccccgcugug cuugcaccug gcauccucgu 240
gcuccuguuu accuuggugc agaggagcaa uggggagugu aaagaggcac uagcaaaguc 300
cgagaugaau gugaauauga aguaucagcu ucccaacuuc accgcggaaa cacccaucca 360
gaaugucauu cuacaugagc aucacauuuu ccuuggugcc acuaacuaca uuuauguuuu 420
aaaugaggaa gaccuucaga agguugcuga guacaagacu gggccugugc uggaacaccc 480
agauuguuuc ccaugucagg acugcagcag caaagccaau uuaucaggag guguuuggaa 540
agauaacauc aacauggcuc uaguugucga caccuacuau gaugaucaac ucauuagcug 600
uggcagcguc aacagaggga ccugccagcg acaugucuuu ccccacaauc auacugcuga 660
cauacagucg gagguucacu gcauauucuc cccacagaua gaagagccca gccagugucc 720
ugacugugug gugagcgccc ugggagccaa aguccuuuca ucuguaaagg accgguucau 780
caacuucuuu guaggcaaua ccauaaauuc uucuuauuuc ccagaucauc cauugcauuc 840
gauaucagug agaaggcuaa aggaaacgaa agaugguuuu auguuuuuga cggaccaguc 900
cuacauugau guuuuaccug aguucagaga uucuuacccc auuaaguaug uccaugccuu 960
ugaaagcaac aauuuuauuu acuucuugac gguccaaagg gaaacucuag augcucagac 1020
uuuucacaca agaauaauca gguucuguuc cauaaacucu ggauugcauu ccuacaugga 1080
aaugccucug gaguguauuc ucacagaaaa gagaaaaaag agauccacaa agaaggaagu 1140
guuuaauaua cuucaggcug cguaugucag caagccuggg gcccagcuug cuagacaaau 1200
aggagccagc cugaaugaug acauucuuuu cgggguguuc gcacaaagca agccagauuc 1260
ugccgaacca auggaucgau cugccaugug ugcauucccu aucaaauaug ucaacgacuu 1320
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ccaaaccauu ucaacugagu uugcuguuaa auugaaaauu gacuuagcca accgagagac 2400
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ccagaucaga auuucacagg auugauugcu gguguugucu caauaucaac agcacuguua 2940
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ccucugacag acaugucccc cauccuaacu aguggggacu cugauauauc caguccauua 3240
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cagcauguag ugauugggcc caguagccug auugugcauu ucaaugaagu cauaggaaga 3360
gggcauuuug guuguguaua ucaugggacu uuguuggaca augauggcaa gaaaauucac 3420
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gagcacuaug uccaugugaa cgcuacuuau gugaacguaa aaugugucgc uccguauccu 4200
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gagcaucaga accagaggcu uggucccaca ggccacggac caauggccug cagccgugac 4500
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agaaaacuca gaagagauag uaaugcucag gacaggagcg gcagccccag aacaggccac 4680
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acugcaaccu ccaccuccca ggcucaagcc ucccgaauag cugggacuac aggcgcacac 4920
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caaaauaaga caaguaauuu guugauaaau auuuuuaaag auaacucagc auguuuguaa 5400
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aggccacaaa aacacugcac ugugaacauu uuagaaaagg uaugucagac ugggauuaau 5880
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gagcuauuac aauccaaaua uugccguuuc auaaauguaa uaaguaauac uaauucacag 6180
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uaccaggguc aagagcauga acgcaucaau agaaagaacu cggggaaaca ucccaucaac 6300
aggacuacac acuuguauau acauucuuga gaacacugca augugaaaau cacguuugcu 6360
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cuaagaauua gauacuuguc acugccuaua ccugcagcug aacugaaugg uacuucguau 6480
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<210> 3
<211> 764
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 3
Met Lys Ala Pro Ala Val Leu Ala Pro Gly Ile Leu Val Leu Leu Phe
1 5 10 15
Thr Leu Val Gln Arg Ser Asn Gly Glu Cys Lys Glu Ala Leu Ala Lys
20 25 30
Ser Glu Met Asn Val Asn Met Lys Tyr Gln Leu Pro Asn Phe Thr Ala
35 40 45
Glu Thr Pro Ile Gln Asn Val Ile Leu His Glu His His Ile Phe Leu
50 55 60
Gly Ala Thr Asn Tyr Ile Tyr Val Leu Asn Glu Glu Asp Leu Gln Lys
65 70 75 80
Val Ala Glu Tyr Lys Thr Gly Pro Val Leu Glu His Pro Asp Cys Phe
85 90 95
Pro Cys Gln Asp Cys Ser Ser Lys Ala Asn Leu Ser Gly Gly Val Trp
100 105 110
Lys Asp Asn Ile Asn Met Ala Leu Val Val Asp Thr Tyr Tyr Asp Asp
115 120 125
Gln Leu Ile Ser Cys Gly Ser Val Asn Arg Gly Thr Cys Gln Arg His
130 135 140
Val Phe Pro His Asn His Thr Ala Asp Ile Gln Ser Glu Val His Cys
145 150 155 160
Ile Phe Ser Pro Gln Ile Glu Glu Pro Ser Gln Cys Pro Asp Cys Val
165 170 175
Val Ser Ala Leu Gly Ala Lys Val Leu Ser Ser Val Lys Asp Arg Phe
180 185 190
Ile Asn Phe Phe Val Gly Asn Thr Ile Asn Ser Ser Tyr Phe Pro Asp
195 200 205
His Pro Leu His Ser Ile Ser Val Arg Arg Leu Lys Glu Thr Lys Asp
210 215 220
Gly Phe Met Phe Leu Thr Asp Gln Ser Tyr Ile Asp Val Leu Pro Glu
225 230 235 240
Phe Arg Asp Ser Tyr Pro Ile Lys Tyr Val His Ala Phe Glu Ser Asn
245 250 255
Asn Phe Ile Tyr Phe Leu Thr Val Gln Arg Glu Thr Leu Asp Ala Gln
260 265 270
Thr Phe His Thr Arg Ile Ile Arg Phe Cys Ser Ile Asn Ser Gly Leu
275 280 285
His Ser Tyr Met Glu Met Pro Leu Glu Cys Ile Leu Thr Glu Lys Arg
290 295 300
Lys Lys Arg Ser Thr Lys Lys Glu Val Phe Asn Ile Leu Gln Ala Ala
305 310 315 320
Tyr Val Ser Lys Pro Gly Ala Gln Leu Ala Arg Gln Ile Gly Ala Ser
325 330 335
Leu Asn Asp Asp Ile Leu Phe Gly Val Phe Ala Gln Ser Lys Pro Asp
340 345 350
Ser Ala Glu Pro Met Asp Arg Ser Ala Met Cys Ala Phe Pro Ile Lys
355 360 365
Tyr Val Asn Asp Phe Phe Asn Lys Ile Val Asn Lys Asn Asn Val Arg
370 375 380
Cys Leu Gln His Phe Tyr Gly Pro Asn His Glu His Cys Phe Asn Arg
385 390 395 400
Thr Leu Leu Arg Asn Ser Ser Gly Cys Glu Ala Arg Arg Asp Glu Tyr
405 410 415
Arg Thr Glu Phe Thr Thr Ala Leu Gln Arg Val Asp Leu Phe Met Gly
420 425 430
Gln Phe Ser Glu Val Leu Leu Thr Ser Ile Ser Thr Phe Ile Lys Gly
435 440 445
Asp Leu Thr Ile Ala Asn Leu Gly Thr Ser Glu Gly Arg Phe Met Gln
450 455 460
Val Val Val Ser Arg Ser Gly Pro Ser Thr Pro His Val Asn Phe Leu
465 470 475 480
Leu Asp Ser His Pro Val Ser Pro Glu Val Ile Val Glu His Thr Leu
485 490 495
Asn Gln Asn Gly Tyr Thr Leu Val Ile Thr Gly Lys Lys Ile Thr Lys
500 505 510
Ile Pro Leu Asn Gly Leu Gly Cys Arg His Phe Gln Ser Cys Ser Gln
515 520 525
Cys Leu Ser Ala Pro Pro Phe Val Gln Cys Gly Trp Cys His Asp Lys
530 535 540
Cys Val Arg Ser Glu Glu Cys Leu Ser Gly Thr Trp Thr Gln Gln Ile
545 550 555 560
Cys Leu Pro Ala Ile Tyr Lys Val Phe Pro Asn Ser Ala Pro Leu Glu
565 570 575
Gly Gly Thr Arg Leu Thr Ile Cys Gly Trp Asp Phe Gly Phe Arg Arg
580 585 590
Asn Asn Lys Phe Asp Leu Lys Lys Thr Arg Val Leu Leu Gly Asn Glu
595 600 605
Ser Cys Thr Leu Thr Leu Ser Glu Ser Thr Met Asn Thr Leu Lys Cys
610 615 620
Thr Val Gly Pro Ala Met Asn Lys His Phe Asn Met Ser Ile Ile Ile
625 630 635 640
Ser Asn Gly His Gly Thr Thr Gln Tyr Ser Thr Phe Ser Tyr Val Asp
645 650 655
Pro Val Ile Thr Ser Ile Ser Pro Lys Tyr Gly Pro Met Ala Gly Gly
660 665 670
Thr Leu Leu Thr Leu Thr Gly Asn Tyr Leu Asn Ser Gly Asn Ser Arg
675 680 685
His Ile Ser Ile Gly Gly Lys Thr Cys Thr Leu Lys Ser Val Ser Asn
690 695 700
Ser Ile Leu Glu Cys Tyr Thr Pro Ala Gln Thr Ile Ser Thr Glu Phe
705 710 715 720
Ala Val Lys Leu Lys Ile Asp Leu Ala Asn Arg Glu Thr Ser Ile Phe
725 730 735
Ser Tyr Arg Glu Asp Pro Ile Val Tyr Glu Ile His Pro Thr Lys Ser
740 745 750
Phe Ile Arg His Val Asn Ile Ala Leu Ile Gln Arg
755 760
<210> 4
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 4
atccaaccaa atcttttatt aggcatg 27
<210> 5
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 5
cttctggaaa agtagctcgg tagtct 26
<210> 6
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 6
caaatctttt attaggcatg tcaacatc 28
<210> 7
<211> 47
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 7
taatacgact cactataggg agcgatgttg acatgcctaa taaaaga 47
<210> 8
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 8
gtgtgtcaaa cagtattctt g 21
<210> 9
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 9
ttgacatgcc taataaaaga tttggttg 28
Claims (1)
1.扩增装置、测序装置、计算装置和输出装置在制备评价胶质瘤预后性的系统中的应用，其特征在于，所述扩增装置含有采集单元和扩增单元，所述采集单元用于采集模板核酸片段和扩增引物，所述扩增单元用于利用所述扩增引物对所述模板核酸片段进行扩增，得到扩增产物；
所述测序装置用于对所述扩增产物进行核酸序列测序，得到扩增产物序列；
所述计算装置包括存储器和处理器，所述存储器中存储有计算机程序，所述处理器被配置为执行所述存储器中存储的计算机程序，以实现如下判别：
若所述扩增产物序列中含有如SEQ ID NO.1所示的cDNA的序列和/或如SEQ ID NO.2所示的mRNA的序列，则判定所述模板核酸片段对应的胶质瘤预后性较差；
所述输出装置用于输出所述计算装置的判定结果；
所述胶质瘤为选自IDH突变全级别胶质瘤、继发性胶质瘤和IDH突变胶质瘤中的任意一种。
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| CN202010318352.7A CN113528656B (zh) | 2020-04-21 | 2020-04-21 | 评价胶质瘤和/或胃腺癌预后性的试剂盒和系统 |
| PCT/CN2021/088478 WO2021213398A1 (zh) | 2020-04-21 | 2021-04-20 | 评价胶质瘤和/或胃腺癌预后性的试剂盒和系统 |
| US17/919,711 US20230143605A1 (en) | 2020-04-21 | 2021-04-20 | Kit and system for evaluating glioma and/or gastric adenocarcinoma prognosis |
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Also Published As
| Publication number | Publication date |
|---|---|
| CN113528656A (zh) | 2021-10-22 |
| WO2021213398A1 (zh) | 2021-10-28 |
| US20230143605A1 (en) | 2023-05-11 |
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