CN111650373A - 一种宫颈癌的外周血tcr标志物及其检测试剂盒和应用 - Google Patents
一种宫颈癌的外周血tcr标志物及其检测试剂盒和应用 Download PDFInfo
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Abstract
本发明公开了一种宫颈癌的外周血TCR标志物及其检测试剂盒和应用。该标志物包括序列为SEQ ID NO.1～100所示的蛋白中的至少一种。本发明基于高通量测序方法，只需要采取少量外周血，提取RNA，通过对样本的处理建立免疫图谱文库，再经过高通量测序和TCR数据分析，首先确定宫颈癌外周血中特征性TCR序列，然后将待测样本测试结果与该特征性TCR序列比对，从而确定是否患有宫颈癌。本发明能够同时比较巨大数量的宫颈癌特异性TCR序列，相比单独检测一种或几种标记物，具有更高的特异性和准确性，提高了诊断效率。
Description
技术领域
本发明属于基因工程技术领域，具体涉及一种宫颈癌的外周血TCR标志物及其检测试剂盒和应用。
背景技术
根据CGCS第26届中国肿瘤防治协会报告，每年新发宫颈癌病例在50万左右(世界范围)，80％以上的病例发生在发展中国家。我国每年新发病例约13万，占全世界新发病例总数的28％。2018年发布的ICO中国HPV和相关疾病报告显示，在中国15-44岁女性中，子宫颈癌的发病率高居恶性肿瘤第三位，以成为女性癌症死亡的第二大原因。中国2018年约有47739名女性因子宫颈癌死亡。且子宫颈癌死亡率呈上升趋势，威胁中国女性生命安全。高危型别HPV持续感染是子宫颈癌的主要致病“元凶”.HPV感染率很高，一项基于美国人群的研究显示，有性行为的男性和女性一生中感染HPV的几率高达85％-90％。大多数感染可以依靠自身的免疫力清除。小部分患者高危型别HPV持续感染，经数十年时间可导致病变并最终进展为子宫颈癌。
中国女性约98％的子宫颈癌由高危型HPV导致，其中69％子宫颈癌有高危型别HPV16/18感染，23％子宫颈癌有高危型别HPV31/33/45/52/58感染,其他高危型别HPV感染致癌率达6％。目前，子宫颈癌的综合防控主要分为三级。以减少HPV感染为一级预防：1.健康教育；2、接种HPV疫苗。对癌前病变进行筛查、诊断和治疗为二级预防(最有效可控的预防阶段)，现有的主要筛查方法主要有：宫颈细胞学涂片、HPV检测、阴道镜检查后的随访管理。三级预防也是最无奈的方式，对子宫颈癌进行治疗。
目前针对卵巢癌的诊断手段主要分为视诊、触诊、细胞学涂片检查、组织学检查、腔镜检查、影像学检查、肿瘤标志物检查。
1、视诊：在充足照明条件下，直接观察外阴和通过阴道窥器观察阴道及宫颈。可直接观察癌浸范围，肿瘤位置、范围、形态、体积及与周围组织的关系，但收视线范围及肿瘤大小影响，有局限性且无法定性。
2、触诊：肿瘤的质地、浸润范围及其与周围的关系等必须通过触诊确定。有些黏膜下及颈管内浸润，触诊比视诊更准确。但依旧受接触面积等人为因素影响，且对肿瘤无法定性。
3、细胞学涂片检查及组织学检查：细胞学涂片检查目前为发现宫颈癌前病变和早期宫颈癌的主要手段，特别是对临床体征不明显的早期病变的诊断。宫颈癌前病变和早期宫颈癌的诊断均应有活体组织学检查证实。大多采用液基细胞学检查法(TCT)，很少给患者带来痛苦，是目前比较有效的防癌方法。由于我国病理医生的缺乏，涂片的阅读准确性很难达到100％。目前也没有数据证实，现有的检查手段能明显降低人群总体上因宫颈癌病死的人数。关键的还是合理的筛查时间间隔且适龄女性一定要记得复查。
4、腔镜检查：对发现宫颈癌前病变、早期宫颈癌、确定病变部位有重要作用，可提高活检的阳性率。接受阴道镜活检均要做颈管刮术。在强光源照射下通过阴道镜直接观察阴道，放大6-20倍，借以观察肉眼看不到的阴道和宫颈的较微小病变。在可疑部分行定位活检，可提高确诊率。由于临近膀胱组织，活检取样是有比较大的难度，因此对临床医生的要求较高，需要临床医生有比较全面的妇产科手术经验。整个检查过程由于属于侵入性检测，患者需要忍受不适而且活检后普遍需要10-14天的时间恢复。此操作对医院的诊疗条件有一定要求，不具备相应条件的需要转诊上级医院。
5、影像学检查
由于解剖部位表浅，绝大多数宫颈癌经妇科检查及细胞病理学检查即可被确诊。在宫颈癌诊断中影像学检查的价值主要是对肿瘤转移、侵犯范围和程度的了解，以指导临床决策并用于疗效评价。
(1)腹盆腔超声：主要用于宫颈局部病变的观察，同时可以观察盆腔及腹膜后区淋巴结转移情况，以及腹盆腔其它脏器的转移情况。但设备的优劣及操作者经验影响诊断的正确率。
(2)盆腔MRI：依照MRI表现提高术前分期的准确率。同时也可观察双侧腹股沟、盆腔及腹膜后区淋巴结转移的情况。
(3)腹盆腔CT：平扫CT观察宫颈局部病变效果不好，尤其是分期较早的病变；增强CT扫描利于宫颈局部病变的显示，但仍有近50％的病变呈等密度，不能清晰显示。
3、血肿瘤标志物检查方法
肿瘤标志物异常升高可以协助诊断、疗效评价、病情监测和治疗后的随访监测，SCC作为宫颈鳞状细胞癌的重要逼走植物，血清SCC水平超过1.5ng/ml被视为异常。但肿瘤标志物只能起到协助诊断的功能，不能作为宫颈癌确诊的依据。
发明内容
针对现有技术中的上述不足，本发明提供一种宫颈癌的外周血TCR标志物及其检测试剂盒和应用，能准确快速的判断待测样本中是否有较高宫颈癌风险患者。
为实现上述目的，本发明解决其技术问题所采用的技术方案是：
一种宫颈癌的外周血TCR标志物，该标志物包括序列为SEQ ID NO.1～100所示的蛋白中的至少一种，具体序列见表1。
表1标志物序列
进一步地，标志物的蛋白序列为SEQ ID NO.1～100所示的序列经取代、缺失和/或替换一个或多个碱基后，能表达相同功能的蛋白。
进一步地，标志物为外周血TCR标志物CDR3序列。
上述标志物在制备治疗宫颈癌的制剂中的应用。
进一步地，制剂中包括含有该标志物的T细胞受体，或能表达产生该标志物的T细胞受体的质粒、病毒载体或核酸片段。
一种用于宫颈癌检测的试剂盒，包括能与上述标志物发生特异性结合的抗体。
一种制剂，包括能与上述标志物发生特异性结合的抗体；所述制剂可用于对宫颈癌进行诊断、预测、检测或筛查。
一种检测宫颈癌的蛋白质芯片，该蛋白质芯片包括基片和点样在基片上特异性抗体，该特异性抗体为能与上述标志物发生特异性结合的抗体。
本发明的原理为：人体内的B淋巴细胞和T淋巴细胞是获得性免疫系统中重要的两类细胞。B细胞通过细胞表面的B细胞受体(BCR)识别抗原，后期BCR在B细胞分化成浆细胞时，表达成抗体，分泌到细胞外。T细胞通过细胞表面的T细胞受体(TCR)识别抗原。BCR和TCR的多样性是建立获得性免疫系统的基础。BCR多样性的理论值是1018，TCR多样性的理论值是1014。BCR与TCR序列中，抗原决定簇3(CDR3)是决定其抗原特异性最重要的部分，因此CDR3的序列被认为可以代表BCR、TCR序列的特性。
在各种疾病中，随着不同的抗原刺激，BCR和TCR的多样性或者表达水平都会发生改变。因此，利用BCR或者TCR高通量测序结果可以追踪疾病的发生、发展。人体内细胞中，衰老蛋白质降解后，其片段会被运输到细胞表面，通过组织相容性抗原II(MCHII)呈递给免疫系统中的T细胞。正常细胞呈递的抗原片段，由于免疫耐受的关系，不会引起免疫反应。一旦当正常细胞出现癌变后，突变的基因表达的异常蛋白，其片段被呈递到细胞表面后，就会引起人体免疫系统产生针对性的免疫反应。因此，分析BCR或TCR的变化，能够检测出肿瘤的发生和发展。
本发明的有益效果为：
1、本发明中，首先利用1300个非宫颈癌的对照组样本、和100个宫颈癌病人的TCR高通量测序数据建立了人工智能分析模型，通过和这些宫颈癌特异性TCR序列对比，可以清楚的判断待测样本中是否有较高宫颈癌风险者。
2、通过高通量测序分析TCR变化可以发现很早期的宫颈癌，利用宫颈癌特有的TCRCDR3序列分析人的免疫系统中的T细胞对宫颈癌的反应，是一种新型的检测方法。
3、本发明通过采用高通量测序技术同时比较数量巨大的特异性TCR序列，比起单独检测一种或几种标记物，具有更高的特异性和准确性。
4、本发明中使用的高通量测序仪器成本低于大型影像学设备，且可向第三方外包，此外，采样、处理的人力成本低于同时检测多种标记物，也低于大量细胞学检测，因此，本发明大大降低了检测成本。
5、本发明只需要采取少量外周血，采样简便、安全。
6、本发明中所述TCR CDR3序列，可用于宫颈癌的免疫治疗。
附图说明
图1为本发明中，对照组CDR3序列及宫颈癌特征序列。横坐标代表某一特定氨基酸组合的CDR3序列被加入对照序列集合或宫颈癌特征序列集合的先后顺序,纵坐标代表该序列在某一样本中重复出现次数CX的对数值；宫颈癌患者的免疫图谱具有多个种类且重复次数较高的宫颈癌特征序列，健康人极少宫颈癌特征序列，而未知受试者的宫颈癌特征比较明显，说明罹患宫颈癌风险较高。
图2为本发明中，针对宫颈癌特征序列集，计算健康人、非肿瘤病人、非宫颈癌肿瘤患者和宫颈癌患者的宫颈癌特征性指数，健康人、非肿瘤病人、非宫颈癌肿瘤患者的宫颈癌特征性指数均与宫颈癌患者具有显著差异，证明了宫颈癌特征序列集的特异性。据此可以判断未知受试者是否罹患宫颈癌。
具体实施方式
下面对本发明的具体实施方式进行描述，以便于本技术领域的技术人员理解本发明，但应该清楚，本发明不限于具体实施方式的范围，对本技术领域的普通技术人员来讲，只要各种变化在所附的权利要求限定和确定的本发明的精神和范围内，这些变化是显而易见的，一切利用本发明构思的发明创造均在保护之列。
实施例1通过免疫图谱分析，获得宫颈癌TCR标志物CDR3序列集
1、采样及免疫图谱分析(方法参考专利ZL201910300069.9)
采集1301名对照组(包括健康人和非肿瘤疾病病人，1300人用于建立模型，1个健康人用于验证)、101名宫颈癌患者(100人用于建立模型，1人用于验证)及1名未知健康状况受试者的外周血(每人10mL)，通过高通量测序得到受试者和对照组的TCR的抗原决定簇3(CDR3)氨基酸序列，保证每个样本的功能性TCR的CDR3序列总数综合不低于30000；
2、对每个样本的TCR的CDR3序列进行随机不放回抽样，使每个样本的CDR3序列数量总和均为30000。对任一特定CDR3序列X，在单样本测序结果中重复出现次数计为CX；
3、通过分析TCR CDR3数据，确定宫颈癌TCR标志物CDR3序列：
a)将1300名用于建立模型的对照组样本的所有CDR3序列归总去重，设为对照序列集；
b)将100名用于建立模型的宫颈癌样本的所有CDR3序列归总去重，再去除所有与对照序列集中包含序列重复的序列，设为宫颈癌特征序列集。作图如附图1A所示，其中横坐标代表某一特定氨基酸组合的CDR3序列被加入对照序列集合或宫颈癌特征序列集合的先后顺序，纵坐标代表该序列在某一样本中重复出现次数CX的对数值。
c)按照同样作图方法，将1名健康人、1名宫颈癌患者和1名健康状况未知受试者的免疫图谱，参照对照序列集合和宫颈癌特征序列集合进行作图，见附图1B-D。从图中可见，宫颈癌患者的免疫图谱中，含有较多种类且较高重复出现次数的宫颈癌特征序列；健康人的免疫图谱中，只有极少量宫颈癌特征序列；而未知健康状况受试者，有高于健康人的宫颈癌特征序列，说明此人有较高风险罹患宫颈癌。
d)将宫颈癌特征序列集中，将所有出现在两个及以上参与建模宫颈癌样本里的CDR3序列，按“所有参与建模宫颈癌样本里该序列单样本中重复出现次数CX的总和×包含该序列的参与建模宫颈癌样本数”从高到低排序，排名前100者即为宫颈癌TCR标志物CDR3序列，具体序列如SEQ ID NO.1～100所示。
实施例2验证宫颈癌TCR标志物CDR3序列集的特异性
1、采样及免疫图谱分析(方法参考专利ZL201910300069.9)
采集250名非宫颈癌的肿瘤患者、30名未知健康状况受试者的外周血(每人10mL)，通过高通量测序得到受试者和对照组的TCR的抗原决定簇3(CDR3)氨基酸序列，保证每个样本的功能性TCR的CDR3序列总数综合不低于30000；对每个样本的TCR的CDR3序列进行随机不放回抽样，使每个样本的CDR3序列数量总和均为30000。
2、从实施例1的对照组中随机挑选100名健康人及45名非肿瘤疾病病人。
3、根据来自实施例1的100名健康人、45名非肿瘤疾病病人、100名宫颈癌患者，以及实施例2新获取的250名非宫颈癌肿瘤患者、30名未知健康状况受试者的免疫图谱，分析其宫颈癌特征性指数。
其中宫颈癌特征性指数定义为：某个样本中，属于宫颈癌特征序列集的所有CDR3序列在该样本内重复出现次数CX的总和。分析结果见下表2及附图2。宫颈癌组与健康人(p＝1.1E-121)、非肿瘤疾病(p＝1.7E-73)、其它肿瘤(p＝7.4E-220)都有显著差异，这证明了宫颈癌特征序列集的特异性。
表2不同样本组的宫颈癌特征性指数
4、分析各组的宫颈癌特征指数(表3)，未知健康状况受试者(检测样本)中，前17人的宫颈癌特征指数高于“其它肿瘤”组的平均值+2×SD(362+2×719＝1800)，此4人具有较高风险罹患宫颈癌；后13人宫颈癌特征指数接近健康人或非肿瘤疾病组平均值，罹患宫颈癌风险低。与临床体检结果对照后，前17人确为宫颈癌患者，而后13人为健康人。此实施例证明了利用宫颈癌特征序列集及宫颈癌特征性指数，预测受试者罹患宫颈癌风险的可行性。
表3宫颈癌特征性指数分析
| 健康人 | 非肿瘤疾病 | 其他肿瘤 | 宫颈癌 | 检测样本 | |
| Mean | 110.08 | 206.36 | 362.37 | 12914.91 | 3243.30 |
| SD | 37.22 | 592.90 | 718.92 | 2334.05 | 3572.74 |
| mean+2SD | 184.52 | 1392.15 | 1800.21 |
综上所述，本发明所述宫颈癌TCR标志物CDR3序列，确实具有显著的宫颈癌特异性，不仅可以用于宫颈癌预测受试者罹患宫颈癌风险，未来还可用于宫颈癌的生物免疫治疗。
序列表
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Ala Ser Gln Gly Phe Thr Ser Gly Gly Asn Asn Glu Gln Phe
1 5 10
<210> 29
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 29
Ala Ser Ser Gln Ala Phe Gly Ala Asp Tyr Tyr Asn Glu Gln Phe
1 5 10 15
<210> 30
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 30
Ala Ser Gly Leu Gly Thr Thr His Gln Pro Gln His
1 5 10
<210> 31
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 31
Ala Thr Gly Leu Ala Gly Gln Tyr Asn Glu Gln Phe
1 5 10
<210> 32
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 32
Ala Ser Arg Ser Gly Glu Gly Phe Asn Glu Gln Phe
1 5 10
<210> 33
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 33
Ala Ser Ser Tyr Glu Ser Leu Ala Gly Ala Val Gly Thr Gln Tyr
1 5 10 15
<210> 34
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 34
Ala Ser Ser Leu Val Pro Asp Ala Tyr Tyr Gly Tyr Thr
1 5 10
<210> 35
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 35
Ala Ser Arg Gly Arg Arg Glu Gly Pro Thr Asp Thr Gln Tyr
1 5 10
<210> 36
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 36
Ala Ser Ser Pro Val Trp Thr Asp Gly Glu Gln Phe
1 5 10
<210> 37
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 37
Ala Ser Lys Ser Gly Thr Ser Gly Ile Pro Ser Asp Thr Gln Tyr
1 5 10 15
<210> 38
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 38
Ala Ser Ser Leu Phe Arg Thr Gly Arg Asp Ile Gln Tyr
1 5 10
<210> 39
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 39
Ala Ser Ser Glu Leu Pro Gly Gln Thr Tyr Asn Glu Gln Phe
1 5 10
<210> 40
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 40
Ala Ser Ser Phe Glu Arg Ser Gly Ser Ser Ser Tyr Glu Gln Tyr
1 5 10 15
<210> 41
<211> 10
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 41
Ala Ser Met Gly Ser His Tyr Gly Tyr Thr
1 5 10
<210> 42
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 42
Ala Ser Ser Met Gly Gly Val Asn Ser Ser Tyr Glu Gln Tyr
1 5 10
<210> 43
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 43
Ala Ser Glu Pro Tyr Ser Gly Gly Thr Asp Thr Gln Tyr
1 5 10
<210> 44
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 44
Ala Ser Ser Pro Gly Asp Arg Gly Ile Tyr Glu Gln Phe
1 5 10
<210> 45
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 45
Ala Ser Ser Arg Thr Gly His Lys Glu Thr Gln Tyr
1 5 10
<210> 46
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 46
Ala Ser Ser Gly Asp Arg Glu Gln Glu Thr Gln Tyr
1 5 10
<210> 47
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 47
Ala Ser Ser Thr Glu Gln Gly Pro Tyr Ser Gln Glu Thr Gln Tyr
1 5 10 15
<210> 48
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 48
Ala Ser Ser Leu Glu Ile Ser Arg Phe Gln Glu Thr Gln Tyr
1 5 10
<210> 49
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 49
Ser Ala Gly Lys Thr Arg Met Asn Thr Glu Ala Phe
1 5 10
<210> 50
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 50
Ala Ser Ser Leu Ser Leu Asp Arg Val Thr Ala Tyr Thr
1 5 10
<210> 51
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 51
Ser Ala Val Ala Gly Gln Gly Asp Tyr Gly Tyr Thr
1 5 10
<210> 52
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 52
Ser Ala Ser Gly Arg Ser Ile Leu Gly Asn Glu Gln Phe
1 5 10
<210> 53
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 53
Ala Ser Ser Leu Ser Glu Gly Ser His Tyr Gly Tyr Thr
1 5 10
<210> 54
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 54
Ala Ser Thr Leu Ala Gly Leu Ala Gly Asn Glu Gln Phe
1 5 10
<210> 55
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 55
Ala Ser Ser Ser Val Gly Ser Ser Ser Tyr Asn Ser Pro Leu His
1 5 10 15
<210> 56
<211> 17
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 56
Ala Ser Ser Phe Pro Pro Gly Leu Ala Gly Val Ser Gln Glu Thr Gln
1 5 10 15
Tyr
<210> 57
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 57
Ala Ser Ser Thr Arg Asp Phe Arg Asn Ser Pro Leu His
1 5 10
<210> 58
<211> 11
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 58
Ala Ser Ser Thr Glu Leu Arg Asn Glu Gln Phe
1 5 10
<210> 59
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 59
Ala Ser Ser Pro Gly Asp Ala Lys Val Asp Glu Gln Tyr
1 5 10
<210> 60
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 60
Ala Ser Thr Pro Phe Ala Gly Gly His Asn Glu Gln Phe
1 5 10
<210> 61
<211> 16
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 61
Ala Ser Ser Ser Thr Thr Ser Arg Gly Arg Arg Ile Tyr Glu Gln Tyr
1 5 10 15
<210> 62
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 62
Gln Gln Pro Gly Pro Arg Pro Val Gly Tyr Ala Val
1 5 10
<210> 63
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 63
Ala Ser Ser Gln Asp Asp Trp Asp Asn Ser Pro Leu His
1 5 10
<210> 64
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 64
Ala Ser Lys Gln Gly Asp Gln Phe
1 5
<210> 65
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 65
Ala Ser Ser Glu Ala Gly Ala Gly Gly Tyr Glu Gln Phe
1 5 10
<210> 66
<211> 11
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 66
Ser Ala Ala Gly Thr Gly Tyr Ser Glu Gln Tyr
1 5 10
<210> 67
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 67
Ala Ser Ser Val Val Ala Gly Gly Arg Gln Glu Thr Gln Tyr
1 5 10
<210> 68
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 68
Ala Ser Ser Pro Gln Gly Tyr Gly Phe Tyr Glu Gln Tyr
1 5 10
<210> 69
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 69
Ala Ser Ser Leu Leu Arg Ala Pro Thr Asp Thr Gln Tyr
1 5 10
<210> 70
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 70
Ala Ser Ser Leu Tyr Asp Arg Arg Gln Pro Gln His
1 5 10
<210> 71
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 71
Ser Ala Val Ser Asp Gly Phe Ser Tyr Glu Gln Tyr
1 5 10
<210> 72
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 72
Ala Ser Ser Pro Gly Pro Gln Pro His Tyr Glu Gln Tyr
1 5 10
<210> 73
<211> 16
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 73
Ala Ser Ser Ser Arg Pro Thr Gly Gly Ala Gly Ala Asn Val Leu Thr
1 5 10 15
<210> 74
<211> 16
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 74
Ala Ser Ser Pro Arg Arg Gln Pro Asn Met Asn Thr Gly Glu Leu Phe
1 5 10 15
<210> 75
<211> 11
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 75
Ser Val Tyr Arg Leu Ser Leu Asn Glu Gln Phe
1 5 10
<210> 76
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 76
Ala Ser Ser Leu Lys Val Thr Ser Asn Glu Gln Phe
1 5 10
<210> 77
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 77
Ala Ser Ser Thr Arg Leu Ala Gly Gly Ile Asp Glu Gln Phe
1 5 10
<210> 78
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 78
Ser Ala Val Thr Ser Gly Ser Pro Asn Tyr Glu Gln Tyr
1 5 10
<210> 79
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 79
Ala Ser Arg Gly Val Gln Ser Gly Tyr Glu Gln Tyr
1 5 10
<210> 80
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 80
Ala Ser Ser Ser Pro Gly Leu Thr Ser Leu Asn Thr Glu Ala Phe
1 5 10 15
<210> 81
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 81
Ala Ser Ser Leu Ala Ser Trp Met Ala Thr Asp Thr Gln Tyr
1 5 10
<210> 82
<211> 10
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 82
Ala Ile Ser Glu Thr Gly Gln Glu Gln His
1 5 10
<210> 83
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 83
Ala Ser Ser Leu Gly Ala Val Val Gly Thr Gln Tyr
1 5 10
<210> 84
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 84
Ala Ser Ser Pro Leu Asp Gly Gly His Leu Asp Glu Gln Tyr
1 5 10
<210> 85
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 85
Ala Thr Lys Gly Ser Arg Trp Tyr
1 5
<210> 86
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 86
Ala Ser Ser Phe Gly Gln Pro Gly Asn Ile Gln Tyr
1 5 10
<210> 87
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 87
Ala Ser Ser Leu Ala Gly Asp Ser Thr Pro Tyr Glu Gln Tyr
1 5 10
<210> 88
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 88
Ala Ser Ser Glu Trp Asp Arg Ile Asp Thr Gln Tyr
1 5 10
<210> 89
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 89
Ala Ser Arg Asp Gln Ala Ser Ser Gly Asn Thr Ile Tyr
1 5 10
<210> 90
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 90
Ser Ala Gly Thr Leu Ala Gly Val Asn Glu Gln Phe
1 5 10
<210> 91
<211> 16
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 91
Ala Ser Ser Tyr Ser Ile Thr Ser Gly Thr Ser Thr Asp Thr Gln Tyr
1 5 10 15
<210> 92
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 92
Ala Ser Ser Ser Asn Leu Ile Arg Pro Pro Tyr Glu Gln Tyr
1 5 10
<210> 93
<211> 11
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 93
Ala Ser Ser Ser Thr Ser Val Asp Glu Gln Tyr
1 5 10
<210> 94
<211> 9
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 94
Ala Val Gln Ser Tyr Ala Glu Gln Phe
1 5
<210> 95
<211> 16
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 95
Ala Ser Leu Ala Gly Leu Ala Gly Gly Ser Ser Tyr Asn Glu Gln Phe
1 5 10 15
<210> 96
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 96
Ala Ile Ser Glu Ser Glu Arg Leu Thr Asp Thr Gln Tyr
1 5 10
<210> 97
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 97
Ala Ser Thr Pro Glu Ala Thr Ser Val Glu Gln Tyr
1 5 10
<210> 98
<211> 11
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 98
Ser Val Glu Asp Arg Tyr Gly Gly Glu Leu Phe
1 5 10
<210> 99
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 99
Ala Ser Ser Glu Leu Phe Pro Ser Gln Thr Gly Thr Pro Leu His
1 5 10 15
<210> 100
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 100
Ala Ser Thr Lys Thr Gly Gly Leu Gly Ser Tyr Asn Glu Gln Phe
1 5 10 15
Claims (7)
1.一种宫颈癌的外周血TCR标志物，其特征在于，所述标志物包括序列为SEQ ID NO.1～100所示的蛋白中的至少一种。
2.根据权利要求1所述的宫颈癌的外周血TCR标志物，其特征在于，所述标志物的蛋白序列为SEQ ID NO.1～100所示的序列经取代、缺失和/或替换一个或多个碱基后，能表达相同功能的蛋白。
3.权利要求1所述的标志物在制备治疗宫颈癌的制剂中的应用。
4.根据权利要求3所述的应用，其特征在于，所述制剂包括含有该标志物的T细胞受体，或能表达产生该标志物的T细胞受体的质粒、病毒载体或核酸片段。
5.一种用于宫颈癌检测的试剂盒，其特征在于，包括能与权利要求1所述标志物发生特异性结合的抗体。
6.一种制剂，其特征在于，包括能与权利要求1所述标志物发生特异性结合的抗体。
7.一种检测宫颈癌的蛋白质芯片，其特征在于，所述蛋白质芯片包括基片和点样在基片上特异性抗体，所述特异性抗体为能与权利要求1所述标志物发生特异性结合的抗体。
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| PCT/CN2021/101678 WO2022012283A1 (zh) | 2020-07-11 | 2021-06-23 | 一种宫颈癌的外周血tcr标志物及其检测试剂盒和应用 |
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| CN113109564A (zh) * | 2021-03-15 | 2021-07-13 | 成都益安博生物技术有限公司 | 一种急性髓细胞性白血病的外周血tcr标志物及其检测试剂盒和应用 |
| CN113567682A (zh) * | 2021-07-23 | 2021-10-29 | 成都益安博生物技术有限公司 | 一种阿尔茨海默病的外周血tcr标志物及其检测试剂盒和应用 |
| WO2022012283A1 (zh) * | 2020-07-11 | 2022-01-20 | 成都益安博生物技术有限公司 | 一种宫颈癌的外周血tcr标志物及其检测试剂盒和应用 |
| WO2022194039A1 (zh) * | 2021-03-15 | 2022-09-22 | 成都益安博生物技术有限公司 | 一种急性b淋巴细胞白血病的外周血tcr标志物及其检测试剂盒和应用 |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2022012283A1 (zh) * | 2020-07-11 | 2022-01-20 | 成都益安博生物技术有限公司 | 一种宫颈癌的外周血tcr标志物及其检测试剂盒和应用 |
| CN113109564A (zh) * | 2021-03-15 | 2021-07-13 | 成都益安博生物技术有限公司 | 一种急性髓细胞性白血病的外周血tcr标志物及其检测试剂盒和应用 |
| WO2022194039A1 (zh) * | 2021-03-15 | 2022-09-22 | 成都益安博生物技术有限公司 | 一种急性b淋巴细胞白血病的外周血tcr标志物及其检测试剂盒和应用 |
| CN113567682A (zh) * | 2021-07-23 | 2021-10-29 | 成都益安博生物技术有限公司 | 一种阿尔茨海默病的外周血tcr标志物及其检测试剂盒和应用 |
| WO2023000688A1 (zh) * | 2021-07-23 | 2023-01-26 | 成都益安博生物技术有限公司 | 一种阿尔茨海默病的外周血tcr标志物及其检测试剂盒和应用 |
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| WO2022012283A1 (zh) | 2022-01-20 |
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