CN106192017A - 一种用于检测高血压的基因芯片及其试剂盒 - Google Patents
一种用于检测高血压的基因芯片及其试剂盒 Download PDFInfo
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Abstract
本发明提供了一种用于检测高血压的基因芯片及其试剂盒，其含有适配子。所述试剂盒能够快速的检测人甘油三磷酸脱氢酶，通过检测所述酶的量，可以用来鉴定样本是否为原发性高血压。本发明的试剂盒采用芯片的方式进行检测，芯片检测具有检测效率高，灵敏度好，应用范围广发的效果，可以大面积推广使用。
Description
技术领域
本发明属于生物领域，具体的说，涉及一种与人甘油三磷酸脱氢酶具有特异性结合的生物芯片，已经相应的制备得到的试剂盒。
背景技术
高血压(hypertension)是一种常见的以动脉血压持续升高为主要表现的慢性疾病，指在静息状态下动脉舒张压>90mmHg。而收缩压在120-139mmHg和/或舒张压在80-89mmHg之间称为高血压前期或正常高值。
高血压是心脑血管病最主要的危险因素，其脑卒中、心肌梗死、心力衰竭及慢性肾脏病等主要并发症，不仅致残、致死率高，而且严重消耗医疗和社会资源，给家庭和国家造成沉重负担。大量长期的临床观察表明高血压前期也是心脑血管疾病和肾病的独立危险因素。我国人群50年来高血压患病率的明显上升趋势。根据2002年调查数据，我国18岁以上成人高血压患病率为18.8％，估计目前我国约有2亿高血压患者，每10个成年人中就有2人患有高血压，约占全球高血压总人数的1/5。高血压前期在我国的发病率约为44％-60％，以中青年为主；其中45％的高血压前期在十年内发展为高血压，是我国高血压患者的"后备军"。国内外的实践证明，高血压是可以预防和控制的疾病，降低高血压患者的血压水平，可明显减少脑卒中及心脏病事件，显著改善患者的生存质量，有效降低疾病负担。
高血压发生的病理学基础是血管重塑造成的血管结构与功能改变，因此高血压的本质是一种血管性病变。高血压导致的靶器官损伤也是以血管损伤为基础的器官功能衰减过程。血管损伤是高血压发生的病理基础，也是高血压维持、发展的结构基础。因此如何采取简便、快捷、准确的手段在高血压早期即筛查出高血压及血管损伤的高危患者，及早采取治疗预防措施，对于提高心血管疾病的控制率、降低心血管并发症的死亡率、改善病人的生活质量、减少国家医疗费用支出等都具有重要意义，也是目前医学研究及临床应用的重要课题。
原发性高血压(essential hypertension，EH)是最常见的心血管疾病，其发病率逐年上升，并可引起严重的心、脑、肾并发症，是脑卒中和冠心病的主要危险因素。EH是一种由遗传因素和环境因素相互作用而发病的多基因病，寻找EH相关基因进而阐明高血压发病的遗传机制已经成为目前研究的热点(HarrapSB.Hypertension：genes versusenvironment.Lancet，1994，344：169-171)。
基因芯片(Gene chip)又称DNA芯片(DNA CHIP)、DNA阵列(DNA arrays)、寡核苷酸微芯片(oligonucleotide micro-chip)，是指将许多特定的寡核苷酸片段或基因片段作为探针，有规律地排列固定于支持物上，然后与待测的标记样品的基因按碱基配对原理进行杂交，再通过检测系统等对芯片进行扫描，并配以计算机系统对杂交信号的强度进行分析和处理，从而迅速得出所要的信息。目前，基因芯片技术正逐渐应用到生命科学各个领域中。
基因芯片制备技术：基因芯片的实质是高度集成的寡核苷酸阵列，制造基因芯片首先要解决的技术是如何在芯片片基上定位合成高密度的核酸探针。目前，基因芯片的制备技术主要有原位合成与合成后点样两种。
基因芯片在临床疾病诊断中的应用：从正常人的基因组中分离出DNA与DNA芯片杂交就可以得出标准图谱。从病人的基因组中分离出DNA与DNA芯片杂交就可以得出病变图谱。这种基因芯片诊断技术以其快速、高效、敏感、经济、平行化、自动化等特点，将成为一项现代化诊断新技术。例如，Affymetrix公司和Oncormed公司合作研制的p53基因芯片可监测50％以上肿瘤患者的p53基因突变，可在癌症早期诊断中发挥作用。基因芯片在感染性疾病、遗传性疾病、重症传染病和恶性肿瘤等疾病的临床诊断方面具有独特的优势。与传统检测方法相比，它可以在一张芯片同时对病人进行多种疾病的检测，勿需机体处于免疫应答反应期，能及早诊断，待测样品用量小；能特异性检测病原微生物的亚型及变异；可帮助医生及患者从“系统、血管、组织和细胞层次(第二阶段医学)”转变到“DNA、RNA、蛋白质及其相互作用层次(第三阶段医学)”上了解疾病的发生、发展过程。这些特点使得医务人员在短时间内，可以掌握大量的疾病诊断信息，有助于医生在短时间内找到正确的治疗措施。
大量研究表明，高血压的发生是遗传和环境因素相互作用所致。筛选与中国人群高血压相关的易感基因片段，对于在人群中开展综合防治项目有重要意义。将遗传上相对易感的人作为高危人群，进行针对性的环境与行为干预，可以在有限的卫生资源条件下，更好地达到预防高血压的目的。本发明所要解决的技术问题是针对目前对原发性高血压疾病的诊断过程中，难以确定的状况，提供一种方便、快捷的辅助检测原发性高血压的诊断芯片。
GPD2基因已经报道是与原发性高血压紧密相关的。现有技术中检测GPD2的表达量主要通过酶联免疫反应进行检测，所述检测比较费时费力，效率低下。如果能筛选出甘油三磷酸脱氢酶的适配子，然后将其包被在生物芯片上，再结合SPR生物传感器进行检测，将非常的快捷和精确。
发明内容
为解决以上技术问题，本发明的目的之一在于提供一种与甘油三磷酸脱氢酶结合力更强、稳定性好、精确性高、重复性好的适配子。
本发明目的之二在于提供一种与人甘油三磷酸脱氢酶GPD2具有特异性的适配子所构成的生物芯片。
本发明目的之三在于提供一种试剂盒，所述试剂盒其含有上述的检测芯片。
本发明目的是这样实现的：一种与人甘油三磷酸脱氢酶GPD2具有特异性的适配子，其特征在于：他具有SEQ ID No.1-SEQ ID No.26的序列之一。
一种由与人甘油三磷酸脱氢酶GPD2具有特异性的适配子构成的的生物芯片，包括玻璃基底(1)，玻璃基底(1)上依次附着有金膜(2)、连接层(3)和表面基质(4)，所述金膜(2)的流池包被有SEQ ID No.1-SEQ ID No.26序列的适配子。
上述金膜⑵在包被适配子前,先预处理，即在1.0mol/L NaOH中浸泡清洗20min，取出后用去离子水清洗3次,然后放入Piranha溶液中处理15min,取出后立即投入无水乙醇中浸泡5min,氮气吹干。
上述适配子包被在金膜(2)上之前，先预处理，即先将适配子溶解到PBS缓冲液，95℃变性3min,迅速冰浴2min,随后巯基法进行固定。
人甘油三磷酸脱氢酶GPD2适配子经过以下方法制备筛选得到：
构建随机单链DNA(ssDNA)文库和引物：构建ssDNA文库，两端为固定序列，中间35个核苷酸为随机序列，其中N代表A，T，C，G中的任意一个：5’-ACTTTAGTAGCCAGTGAGAC-N35-GGAGCAGTACAGTGATCAGT-3’。
上游引物为5，-ACTTTAGTAGCCAGTGAGAC-3’，下游引物为5’-ACTGATCACTGTACTGCTCC-3’。下游引物序列5’端需用生物素标记。随机单链DNA文库和引物可由引物合成公司合成。
双链DNA(dsDNA)文库的PCR扩增及回收；
单链DNA文库的PCR扩增及回收；
采用SELEX筛选，将纯化后的目的基因PCR产物与克隆载体pMD18-Tsimplevector连接，测序。
适配子包被生物芯片
我们在表面基质上包被亲和性最强的适配子，然后结合SPR生物传感器进行检测。
有益效果：本发明提供了一种试剂盒，所述试剂盒能够快速的检测人甘油三磷酸脱氢酶，通过检测所述酶的量，可以用来鉴定样本是否为原发性高血压。本发明的试剂盒采用芯片的方式进行检测，芯片检测具有检测效率高，灵敏度好，应用范围广发的效果，可以大面积推广使用。
具体实施方式：
实施例1适配子的获得
重组人甘油三磷酸脱氢酶-2为市场上购买自ProSpec，产品货号ENZ-437。
构建随机单链DNA(ssDNA)文库和引物：构建ssDNA文库，两端为固定序列，中间35个核苷酸为随机序列，其中N代表A，T，C，G中的任意一个：5’-ACTTTAGTAGCCAGTGAGAC-N35-GGAGCAGTACAGTGATCAGT-3’。
上游引物1为5，-ACTTTAGTAGCCAGTGAGAC-3’，下游引物2为5’-ACTGATCACTGTACTGCTCC-3’。下游引物序列5’端需用生物素标记。随机单链DNA文库和引物可由引物合成公司合成。
(1)利用引物1、引物2扩增单链DNA文库：采用不对称PCR，引物1/引物2浓度比100：1，扩增条件为：94℃预变性:3min，然后94℃变性30s，65℃退火45s，72℃延伸lmin，循环35次，最后72℃延伸7min。将获得的产物(主要为ssDNA)于95℃作用3min，于冰中2min，室温放置l0min，即为ssDNA文库。
(2)反筛与筛选
1OOpmol随机ssDNA文库和5nmol(tRNA+鲑鱼精DNA)加入10μl BSA反筛，在100μ1结合缓冲液中室温孵育1h；然后转移液体，与人甘油三磷酸脱氢酶25℃结合lh，用冲洗缓冲液洗6次，洗去未结合的ssDNA，再加洗脱缓冲液于80℃作用lOmin，洗脱下与人甘油三磷酸脱氢酶结合的ssDNA，经酚-氯仿抽提、乙醇沉淀。将ssDNA溶解于20μ1TE缓冲液中，用作扩增的模板，进行下一轮筛选，重复筛选20轮。
(3)测定亲和性
将第20轮得到的ssDNA文库经过不对称PCR扩增获得生物素标记的ssDNA文库，将纯化获得的人甘油三磷酸脱氢酶GPD2蛋白用碳酸盐(pH9.6)缓冲液稀释至10μg/ml包被酶联板，4℃过夜，PBST(PBS+Tween)洗涤3次，3min/次；3％BSA37℃封闭1小时，PBST洗涤3次,3min/次；用SELEX binding buffer稀释的第15轮生物素标记的ssDNA 0.05μg/孔，同时加入不同浓度梯度的人甘油三磷酸脱氢酶GPD2蛋白，37℃温育60min，PBST洗涤4次，3min/次；1：2000稀释的链霉亲和素标记的辣根过氧化物酶37℃孵育30min，PBST洗涤4次，3min/次；加入四甲基联苯胺(tetramethylbenzidine,TMB)显色液37℃显色15min；2mol/L浓硫酸终止反应，酶标仪于450nm处测定A值。A值为0.160.回收产物连接，20轮后，将筛选产物连接到PMD18-T simple vector(TaKaRa公司)。参考TaKaRa公司pMD18_Tsimple vector的产品说明，将纯化后的目的基因PCR产物与克隆载体pMD18-Tsimplevector连接，目的基因片段与载体片段反应体系如下：
pMD18-T/PCR产物/连接缓冲液：1μl/9μ1/10μl 16℃连接过夜，连接产物的转化：
A、将目的基因片段与pMD18-T simple vector的连接产物20μl加入含有100μI的E.coli DH5a感受态细胞的EP管中，冰浴30min
B、放入42℃水浴热休克lmin，快速将管取出冰浴2min
C、每管加SOC培养液600μl，37℃摇床，150rpm，培养60min，使细菌复苏并表达质
粒编码的抗生素性标记基因。
D、将适当体积已转化的感受态细胞涂布于含有氨苄青霉素100μg/ml的LB平板上，将平板置于室温直至液体被吸收。
E、倒置平板，于37℃恒温培养，12-16小时后出现菌落，随机挑取几个菌落PCR鉴定。
F、随机挑取110个单克隆加入盛有600μl LB液体培养基(含氨苄青霉素100μg/ml)的1.5ml EP管中，37℃摇床，200rpm，培养过夜。次日加入600μl(等量)80％高压灭菌甘油，密封管口，于-70摄氏度保存菌种。
重组质粒的提取，采用质粒提取试剂盒快速提取。
PCR鉴定：
以提取质粒为模板，加入引物1与引物2进行PCR扩增反应，产物电泳。
适体亲和性检测
将挑取的单个克隆经过不对称PCR扩增后得到ssDNA，将此ssDNA与包被好的1μg/孔蛋白结合，酶联法测定其亲和性。
亲和性如下表所示。
测序
挑取单克隆送测序公司测序
所述适配子的序列如SEQ ID NO：1-26所示。
采用本领域常用的Kd测量方法，测得本申请26个序列的Kd解离常数如下：
实施例3适配子包被生物芯片
我们在表面基质上包被亲和性最强的适配子SEQ ID NO：1-26，然后结合SPR生物传感器进行检测，具体方法如下：
生物芯片表面预处理将表面基质镀有链霉亲和素的金膜电极的生物芯片放入1.0mol/LNaOH中浸泡清洗20min，取出后用去离子水清洗3次，然后将生物芯片放入Piranha溶液中处理15min,取出后立即将其投入无水乙醇中浸泡5min,氮气吹干备用。
适配子预处理：PBS缓冲液溶解，95℃变性3min，迅速冰浴2min，随后巯基法进行固定。
巯基法探针固定：经过变性处理的生物素化的适配子覆盖经预处理的芯片表面基质，使适配子牢固的固定在生物芯片上，适配子采用最佳浓度1.0umol/L,2h后PBS缓冲液清洗3次，0.02％BSA封闭约lh，PBS缓冲液清洗。
生物芯片制作完成后，再将包被好适配子的生物芯片载入SPR传感器中，通入过量的蛋白，使其与芯片上的适配子进行反应，最后记录实验结果如下：
根据检测结果，我们可以得出流池1(阴性对照)的角度差为-5.8，流池2-27(检测流池)的角度差依次为：290.5、289.6、278.6、284.9、297.6、287.7、284.0、286.3、286.0、284.1、288.9、284.8、279.6、274.0、290.3、279.1、277.4、286.5、277.5、280.2、278.7、284.3、288.6、289.6、287.5、286.6。两者间有显著差异(P<O.01)。说明通过SPR生物传感器能够对人人甘油三磷酸脱氢酶GPD2进行定量的分析。
实施例4所述适配子特异性分析以及稳定性分析
分别采用NADH脱氢酶、人Ⅲ型乙醇脱氢酶、BSA，人2型11β-羟基类固醇脱氢酶蛋白与26条适配子进行特异性检测，经过结合试验发现，这些适配子都不与这些蛋白相结合，而只与人甘油三磷酸脱氢酶结合保持结合活性。
将所述的适配子，取0.5ug，分别置于常温的血清、水溶液中，放置四周。通过RT-PCR检测，发现四周的放置其结构稳定，没有被降解。说明适配子具有较强的稳定性。
序列表
〈110〉朱继平
〈120〉一种用于检测高血压的基因芯片及其试剂盒
〈160〉26
〈210〉1
〈211〉75
〈212〉DNA
〈213〉人工序列
〈400〉GPD2-1
ACTTTAGTAGCCAGTGAGACCAATTAATACTAATACCATAATACTTCCTTCTCTTGGAGCAGTACAGTGATCAGT
〈210〉2
〈211〉75
〈212〉DNA
〈213〉人工序列
〈400〉GPD2-2
ACTTTAGTAGCCAGTGAGACCTTCTTTCTATACCCTTTAATTCTCCCTTTAACATGGAGCAGTACAGTGATCAGT
〈210〉3
〈211〉75
〈212〉DNA
〈213〉人工序列
〈400〉GPD2-3
ACTTTAGTAGCCAGTGAGACTCTATCATACCATATACATAAAAACTATTCATACTGGAGCAGTACAGTGATCAGT
〈210〉4
〈211〉75
〈212〉DNA
〈213〉人工序列
〈400〉GPD2-4
ACTTTAGTAGCCAGTGAGACTAATATTATATCCCTTCTTCATAATCATCCCTTTAGGAGCAGTACAGTGATCAGT
〈210〉5
〈211〉75
〈212〉DNA
〈213〉人工序列
〈400〉GPD2-5
ACTTTAGTAGCCAGTGAGACTCTCAATACCATTTATACATTATACCTCTTAAACAGGAGCAGTACAGTGATCAGT
〈210〉6
〈211〉75
〈212〉DNA
〈213〉人工序列
〈400〉GPD2-6
ACTTTAGTAGCCAGTGAGACAACCTCTACAAATATTATCTCTATTTATAATCCCTGGAGCAGTACAGTGATCAGT
〈210〉7
〈211〉75
〈212〉DNA
〈213〉人工序列
〈400〉GPD2-7
ACTTTAGTAGCCAGTGAGACTACCAATTTAATCTAAAACCCATATTTTCCACCACGGAGCAGTACAGTGATCAGT
〈210〉8
〈211〉75
〈212〉DNA
〈213〉人工序列
〈400〉GPD2-8
ACTTTAGTAGCCAGTGAGACACACATTCCCACCTAACTTCTTTAACTTAAATACTGGAGCAGTACAGTGATCAGT
〈210〉9
〈211〉75
〈212〉DNA
〈213〉人工序列
〈400〉GPD2-9
ACTTTAGTAGCCAGTGAGACATTTCCCTACTTATTCTCATCATCACTCCACCCTAGGAGCAGTACAGTGATCAGT
〈210〉10
〈211〉75
〈212〉DNA
〈213〉人工序列
〈400〉GPD2-10
ACTTTAGTAGCCAGTGAGACTACAATATCTTTTATCTTAATACACTTACATTACCGGAGCAGTACAGTGATCAGT
〈210〉11
〈211〉75
〈212〉DNA
〈213〉人工序列
〈400〉GPD2-11
ACTTTAGTAGCCAGTGAGACCATTATATAAACCTTAACTCAACTTCTTCCCCATAGGAGCAGTACAGTGATCAGT
〈210〉12
〈211〉75
〈212〉DNA
〈213〉人工序列
〈400〉GPD2-12
ACTTTAGTAGCCAGTGAGACATCCCATACCATCCATAACCTCCATACCTCAAAACGGAGCAGTACAGTGATCAGT
〈210〉13
〈211〉75
〈212〉DNA
〈213〉人工序列
〈400〉GPD2-13
ACTTTAGTAGCCAGTGAGACATCATTATTTTTCTATAATTCTATTATATCCACTAGGAGCAGTACAGTGATCAGT
〈210〉14
〈211〉75
〈212〉DNA
〈213〉人工序列
〈400〉GPD2-14
ACTTTAGTAGCCAGTGAGACCACCCCACAAAATCTTACTCCACATTCAATCACTAGGAGCAGTACAGTGATCAGT
〈210〉15
〈211〉75
〈212〉DNA
〈213〉人工序列
〈400〉GPD2-15
ACTTTAGTAGCCAGTGAGACCAAATCATCTACTCTTCTCTCCAACTTCTACATAAGGAGCAGTACAGTGATCAGT
〈210〉16
〈211〉75
〈212〉DNA
〈213〉人工序列
〈400〉GPD2-16
ACTTTAGTAGCCAGTGAGACCTATTACTTTTCTACCAACTCCTTTTCCTTCTTCAGGAGCAGTACAGTGATCAGT
〈210〉17
〈211〉75
〈212〉DNA
〈213〉人工序列
〈400〉GPD2-17
ACTTTAGTAGCCAGTGAGACCAAAATCACCAACACAAACAATCTTTTTCCTTATCGGAGCAGTACAGTGATCAGT
〈210〉18
〈211〉75
〈212〉DNA
〈213〉人工序列
〈400〉GPD2-18
ACTTTAGTAGCCAGTGAGACTCCACTCTCTCAACTCTAACTACAACTCCTCTCTAGGAGCAGTACAGTGATCAGT
〈210〉19
〈211〉75
〈212〉DNA
〈213〉人工序列
〈400〉GPD2-19
ACTTTAGTAGCCAGTGAGACCATTCATTACCCTACCACATACCATACACCCCTTTGGAGCAGTACAGTGATCAGT
〈210〉20
〈211〉75
〈212〉DNA
〈213〉人工序列
〈400〉GPD2-20
ACTTTAGTAGCCAGTGAGACACTTACCCACTTTCTTTTAATAAACAATACCTCCCGGAGCAGTACAGTGATCAGT
〈210〉21
〈211〉75
〈212〉DNA
〈213〉人工序列
〈400〉GPD2-21
ACTTTAGTAGCCAGTGAGACTCATCTTATCATAAACTTTACATACCAAACATATCGGAGCAGTACAGTGATCAGT
〈210〉22
〈211〉75
〈212〉DNA
〈213〉人工序列
〈400〉GPD2-22
ACTTTAGTAGCCAGTGAGACTCAAAATACCAAAACTTATCACCCTCAACTTACATGGAGCAGTACAGTGATCAGT
〈210〉23
〈211〉75
〈212〉DNA
〈213〉人工序列
〈400〉GPD2-23
ACTTTAGTAGCCAGTGAGACCACCTCACTCCTTATTCCCAACTATTTATAATATAGGAGCAGTACAGTGATCAGT
〈210〉24
〈211〉75
〈212〉DNA
〈213〉人工序列
〈400〉GPD2-24
ACTTTAGTAGCCAGTGAGACCCCTATCCAAACAATCTATCTCCCACCTATCCCCTGGAGCAGTACAGTGATCAGT
〈210〉25
〈211〉75
〈212〉DNA
〈213〉人工序列
〈400〉GPD2-25
ACTTTAGTAGCCAGTGAGACTCTAAATAAACCACCCCAACTTTATATATTACTTCGGAGCAGTACAGTGATCAGT
〈210〉26
〈211〉75
〈212〉DNA
〈213〉人工序列
〈400〉GPD2-26
ACTTTAGTAGCCAGTGAGACCACTTTAACCACAAAACTTCATCCAATACTCAACCGGAGCAGTACAGTGATCAGT
Claims (4)
1.一种基因芯片，其含有适配子。
2.权利要求1所述的基因芯片，其含有适配子，其特征在于：适配子序列为SEQ ID No.1-SEQ ID No. 26的序列之一。
3.一种试剂盒，其中含有权利要求1-2任一项所述的基因芯片。
4.根据权利要求2所述基因芯片或权利要求3所述的试剂盒用于制备高血压诊断试剂中的应用。
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| CN110907643A (zh) * | 2019-12-02 | 2020-03-24 | 中国科学院重庆绿色智能技术研究院 | 一种大肠杆菌检测芯片的制备方法及检测芯片 |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20060110751A1 (en) * | 2004-11-19 | 2006-05-25 | Oy Jurilab Ltd | Method and kit for detecting a risk of essential arterial hypertension |
| CN1861805A (zh) * | 2005-05-11 | 2006-11-15 | 上海市高血压研究所 | 甘油三磷酸脱氢酶基因与原发性高血压的相关性 |
| CN102286607A (zh) * | 2011-08-01 | 2011-12-21 | 四川农业大学 | 一种甘油三磷酸脱氢酶活性检测试剂盒 |
-
2016
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20060110751A1 (en) * | 2004-11-19 | 2006-05-25 | Oy Jurilab Ltd | Method and kit for detecting a risk of essential arterial hypertension |
| CN1861805A (zh) * | 2005-05-11 | 2006-11-15 | 上海市高血压研究所 | 甘油三磷酸脱氢酶基因与原发性高血压的相关性 |
| CN102286607A (zh) * | 2011-08-01 | 2011-12-21 | 四川农业大学 | 一种甘油三磷酸脱氢酶活性检测试剂盒 |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110907643A (zh) * | 2019-12-02 | 2020-03-24 | 中国科学院重庆绿色智能技术研究院 | 一种大肠杆菌检测芯片的制备方法及检测芯片 |
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