12项肿瘤标志物蛋白芯片检测消化系统肿瘤的临床意义

12项肿瘤标志物蛋白芯片检测消化系统肿瘤的临床意义 12项肿瘤标志物蛋白芯片检测消化系 统肿瘤的临床意义 中华首席医学网 2005年09月05日 21:42:47 Monday 622 作者:梁茱 王海枫 吴爱祝 符生苗 《中国热带医学》2005年3月5卷3期 论著 加入收藏夹 【关键词】 肿瘤标志物 摘要:目的 探讨多肿瘤标志物蛋白芯片检测在消化系统肿瘤诊断中的应用价值。 方法 采用蛋白芯片技术检测了消化系统肿瘤患者373例、消化系统良性病变患者105例、健康体检者122例血清中的12种肿瘤标志物(CA199、NSE、CEA、CA242、Ferrtin、β-HCG、AFP、f-PSA、PSA、CA125、HGH、CA153)的水平。 结果 消化系统恶性肿瘤组的阳性率为62.5%(233/373)，其中肝癌阳性率为82.1%(101/123)，胃癌阳性率为47.9%(46/96)，直肠癌阳性率为50.7%(37/73)，结肠癌阳性率为49%(25/51)，胰腺癌阳性率为100%(19/19)，食管癌阳性率为28.6%(2/7)，肛管癌阳性率为75%(3/4)，显著高于良性病变组14.3%(15/105)和健康对照组4.9%(6/122)，而且，肿瘤标志物的联合检测对消化系统恶性肿瘤的检出率明显高于单一标志物的检测。 结论 多肿瘤蛋白芯片检测可以显著提高消化系统恶性肿瘤诊断的敏感性，具有一定的临床应用价值。 关键词:肿瘤标志物;蛋白芯片;消化系统肿瘤 Clinical value of12tumor markers protein biochip in detection of digestive system neoplasm. for computer software engineering GB/T50314-2000 of electrical installations code for construction and acceptance of GBJ232- 92 intelligent building weak design construction figure set GJBT-471 2. system description this times engineering weak design including: hours a Kok information facilities system integrated wiring system (containing room network, communications part) wired TV system information guide and the released system hours II Kok public security system fire automatically alarm system (containing public broadcast part) security technology prevention system security monitoring system out entrance control system car library management system hours three Kok building equipment monitoring system hours four Kok information application system queued station ICU visitation system operation the system monitoring and training system call intercom system hours, five hours Kok Kok intelligent integrated system of information system (1) cabling systems (including computer room network, communication component) computer network systems and voice communications systems Center of information on medical floor computer room located in the engine room. Voice communication system using a local telecommunications operator providing voice service, configuration-independent integrated services digital program-controlled switchboards. Operators providing communications ... If any compatibility requirements, by way of increased access to the Communications Board within the chassis to meet LIANG Zhu，WANG Hai-feng，WU Ai-zhu，et al.(Department of Central Laboratory of Hainan Provincial People's Hospital，Haikou570311，Hainan，P.R.China) Abstract:Objective To discuss the application value of multi-tumor markers protein biochip in digestive system neoplasm. Methods 12Tumor Markers(CA199、NSE、CEA、CA242、 Ferrtin、β-HCG、AFP、f-PSA、PSA、CA125、HGH、CA153)of373pa-tients of digestive system neoplasm，105benign tumor patients of digestive system and122healthy persons were detected by using the technology of Protein Chip. Results The positive rate of the group of digestive system neoplasm is62.5%(233/373)，thereinto，the positive rate of hepatoma is82.1%(101/123)，the positive rate of gastric carcinoma is47.9%(46/96)，the positive rate of rec-tal carcinoma is50.7%(37/73)，the positive rate of colon cancer is49%(25/51)， the positive rate of adenocarcinoma of pancreas is100%(19/19)，the positive rate of carcinoma of esophagus is28.6%(2/7)，the positive rate of carcinoma of anal canal is75%(3/4)，and is obviously higher than the positive rate of the group of benign tumor(14.3%， 15/105)and healthy persons(4.9%，6/122).The rate of combining detection of tumor markers in the digestive system neoplasm is obviously higher than that of the detection of single marker. Conclusion Multi-tumor markers protein biochip could notably improve the sensitivity of diagnosis of the diges-tive system neoplasm，and it is of certain clinical application value. Key words:Tumor marker;Protein chip;Digestive system neoplasm 肿瘤标志物单指标检测在临床上已应用多年，但存在检出率低及费用高的缺点，蛋白芯片是一种集微 电子、微机械、化学物理技术、计算机技术为一体，过程连续化、集成化、微型化的蛋白分析技术， 并具有高速度、高通量、高灵敏度等特性 ,1, 。本实验应用该技术检测了消化系统恶性肿瘤患者、消化 系统良性病变、健康对照者血清中12种肿瘤标志物的水平，以期探讨其对消化系统恶性肿瘤的诊断价值。 1 对象与方法 1.1 研究对象 消化系统恶性肿瘤组为海南省人民医院2004年5,12月的住院患者，均经病理检查 确诊。其中肝癌123例，胃癌96例，直肠癌73例，结肠癌51例，胰腺癌19例，食管癌7例，肛管癌 4例。男 性241例，女性132例，平均年龄59.4岁;消化系统良 性病变组为同期住院患者，其中 for computer software engineering GB/T50314-2000 of electrical installations code for construction and acceptance of GBJ232- 92 intelligent building weak design construction figure set GJBT-471 2. system description this times engineering weak design including: hours a Kok information facilities system integrated wiring system (containing room network, communications part) wired TV system information guide and the released system hours II Kok public security system fire automatically alarm system (containing public broadcast part) security technology prevention system security monitoring system out entrance control system car library management system hours three Kok building equipment monitoring system hours four Kok information application system queued station ICU visitation system operation the system monitoring and training system call intercom system hours, five hours Kok Kok intelligent integrated system of information system (1) cabling systems (including computer room network, communication component) computer network systems and voice communications systems Center of information on medical floor computer room located in the engine room. Voice communication system using a local telecommunications operator providing voice service, configuration-independent integrated services digital program-controlled switchboards. Operators providing communications ... If any compatibility requirements, by way of increased access to the Communications Board within the chassis to meet 肝硬化8例，乙肝7例，肝囊肿5例，胃肠炎54例，胃溃疡7例，胃肠息肉4例，消化道出血11例，胆囊炎11例，胰腺炎3例。男性63例，女性42例，平均年龄57.9岁。健康对照组为2004年11月,12月的体检者，体检中尚未发现任何肿瘤，男性84例，女性38例，平均年龄46.7岁。 1.2 检测方法 1.2.1 血清标本收集 应用无热源和无内毒素的试管抽取晨空腹静脉血2ml，2000,2500g离心8min，采用无溶血和无高脂血清标本。血清分装后置4?C保存，5d内检测，实验前平衡至室温。 1.2.2 仪器与试剂 HD-2001A型系列生物芯片检测仪，由上海数康生物科技有限公司研制生产。试剂盒也是上海数康生物科技有限公司提供的配套试剂，批号为J20040404,20041201。 1.2.3 检测原理 基于双抗体夹心法的化学发光检测方法。在芯片膜上包被12种肿瘤标志物的抗体，捕捉被检者血清中对应的肿瘤标志物，结合标有示踪物的二抗，然后经酶促催化化学反应产生光信号，用专门的芯片检测仪读取光信号，通过软件进行信号值和浓度值的换算，对肿瘤标志物进行定量分析。 1.2.4 操作步骤 按试剂盒操作说明进行。 1.2.5 结果判定 12种肿瘤标志物的临界值为 CA199<35U/L;NSE<13ng/ml;CEA<5ng/ml;CA242<20KU/L;Ferreting<219ng/ml(男性<322ng/ml);β-HCG<3ng/ml;AFP<20ng/ml;f-PSA<1ng/ml;PSA<5ng/ml;CA125<35KU/L; HGH<7.5ng/ml;CA153<35KU/L。超过临界值计算机自动判定为阳性(Fer单独超过临界值不计入阳性数)。 1.2.6 统计学方法 使用SAS6.12统计软件进行统计分析，显著性检验采用χ 2 检验(P<0.01)。 2 结果 2.1 各组阳性率比较 由1可见肿瘤组的阳性率 为62.5%，良性病变组为14.3%，健康对照组为4.9%，肿瘤组阳性率显著高于良性病变组和对照组(χ 2 =76.19，P<0.01;χ 2 =121.93，P<0.01)。特异性为90.7%，准确率为73.2%。 表1 各组阳性率比较(略) for computer software engineering GB/T50314-2000 of electrical installations code for construction and acceptance of GBJ232- 92 intelligent building weak design construction figure set GJBT-471 2. system description this times engineering weak design including: hours a Kok information facilities system integrated wiring system (containing room network, communications part) wired TV system information guide and the released system hours II Kok public security system fire automatically alarm system (containing public broadcast part) security technology prevention system security monitoring system out entrance control system car library management system hours three Kok building equipment monitoring system hours four Kok information application system queued station ICU visitation system operation the system monitoring and training system call intercom system hours, five hours Kok Kok intelligent integrated system of information system (1) cabling systems (including computer room network, communication component) computer network systems and voice communications systems Center of information on medical floor computer room located in the engine room. Voice communication system using a local telecommunications operator providing voice service, configuration-independent integrated services digital program-controlled switchboards. Operators providing communications ... If any compatibility requirements, by way of increased access to the Communications Board within the chassis to meet 2.2 12种肿瘤标志物在各组中的表达 由表2可见肝癌阳性率为82.1%，胰腺癌的阳性率为100.0%，胃癌、直肠癌、结肠癌、食管癌、肛管癌的阳性率分别为47.9%、50.7%、49.0%、28.6%、75.0%。除食管癌外，其余的联合检测阳性率均明显高于单项指标的阳性率。 表2 12种肿瘤标志物在各组中的表达(略) 对照组Control122 6 0.8 0 0.8 0 0 0.8 0 0 0 0.8 1.6 0 4.9 2.3 12种肿瘤标志物在各组中的比较 由表3经χ 2 检验，CA199、CEA、CA242、Fer、β-HCG、AFP、CA125在 肿瘤组和良性组中有显著差异(P<0.01)，而NSE、f-PSA、PSA、HGH、CA153无显著差异(P>0.05)。 表3 12种肿瘤标志物在各组中的比较(略) 3 讨论 消化系统恶性肿瘤严重危害人体的健康，在我国胃癌和肝癌的发病率分别在男性恶性肿瘤中居第一和第三位，而其早诊断、早治疗是提高疗效的最佳手段，但它们的早期共同特点是无明显临床表现，致使诊断时往往已届中晚期，错过了最佳治疗时间，疗效很差，因此，寻找一种高灵敏度、高特异性的无创检查一直为科研和临床所关注。肿瘤标志物的问世带来了惊喜，但绝对理想的肿瘤标志物并不存在，单指标的 AFP是公认的肝癌早期诊断的最佳指标，在本检测始终存在着特异性不强、阳性率较低等不足 ,2, ， 文中其阳性率为47.9%，在以往单指标检测的报导中大致均为60%,70% ,3, ，而徐志宁等报导应用蛋白芯片法将肝癌肿瘤标志物测定的灵敏度和特异性分别提高到了92.5%和92% ,4, ，本实验应用蛋白芯片多指标联合检测其灵敏度和特异性达到82.1%和90.7%，其检出率明显高于AFP单指标的检测率(P<0.01)。从表2可见除食管癌外，其它消化系统恶性肿瘤多指标联合检测可大大提高其检出率，其中以CA199、CEA、CA242、Fer、β-HCG、AFP、CA125的表达显著。 蛋白芯片是近几年快速发展起来的集高通量、高灵敏 度、高特异性、微型化和自动化于一身的高科技方法，克服了单一指标给肿瘤的辅助诊断带来的缺点，并提高了检测的灵敏度和特异性。但是，在应用过程中还存在一些缺陷，如NSE、CA153的检出率低，这可能是由于该蛋白本身的构象问题，导致蛋白质失去活性所致。肿瘤的发生是多步骤、多基因的癌变过程，而且肿瘤细胞生物性的复杂和多肽性，人们还不能单用肿瘤标志物对人体肿瘤做出准确的诊断，但在恶性肿瘤中肿瘤标志物量明显增多。蛋白芯片检测系统检测肿瘤标志物较传统方法快速方便，而且可以同时获得多种肿瘤标志物的信息，更适用于肿瘤高危人群的筛查和肿瘤病人的动态观察。 for computer software engineering GB/T50314-2000 of electrical installations code for construction and acceptance of GBJ232- 92 intelligent building weak design construction figure set GJBT-471 2. system description this times engineering weak design including: hours a Kok information facilities system integrated wiring system (containing room network, communications part) wired TV system information guide and the released system hours II Kok public security system fire automatically alarm system (containing public broadcast part) security technology prevention system security monitoring system out entrance control system car library management system hours three Kok building equipment monitoring system hours four Kok information application system queued station ICU visitation system operation the system monitoring and training system call intercom system hours, five hours Kok Kok intelligent integrated system of information system (1) cabling systems (including computer room network, communication component) computer network systems and voice communications systems Center of information on medical floor computer room located in the engine room. Voice communication system using a local telecommunications operator providing voice service, configuration-independent integrated services digital program-controlled switchboards. Operators providing communications ... If any compatibility requirements, by way of increased access to the Communications Board within the chassis to meet 参考文献: ,1,Sanders G.H.W，ManzA.Chip-based Microsystems for genomic and pro-teomic analysis,J,.Trends Anal chem.，2000，19(6):364,378. ,2,Kayaba H.Tumor Markers:essential diagnostic tools for radiologists,J,.Nippon Igaku Hoshasen Gakkai Zasshi，2003，63(4):133,139. ,3,张军一，左强，廖旺军，等.多肿瘤标志物蛋白芯片检测系统对原发性肝癌的诊断价值,J,.肿 瘤，2004，24(3):254,256. ,4,徐宁志，董志伟.肿瘤的分子生物学基础及其应用,J,.国外医学.肿瘤学分册，2003，30(1):3, 6. 作者单位:海南省人民医院中心实验室，海南海口 570311. for computer software engineering GB/T50314-2000 of electrical installations code for construction and acceptance of GBJ232- 92 intelligent building weak design construction figure set GJBT-471 2. system description this times engineering weak design including: hours a Kok information facilities system integrated wiring system (containing room network, communications part) wired TV system information guide and the released system hours II Kok public security system fire automatically alarm system (containing public broadcast part) security technology prevention system security monitoring system out entrance control system car library management system hours three Kok building equipment monitoring system hours four Kok information application system queued station ICU visitation system operation the system monitoring and training system call intercom system hours, five hours Kok Kok intelligent integrated system of information system (1) cabling systems (including computer room network, communication component) computer network systems and voice communications systems Center of information on medical floor computer room located in the engine room. Voice communication system using a local telecommunications operator providing voice service, configuration-independent integrated services digital program-controlled switchboards. Operators providing communications ... If any compatibility requirements, by way of increased access to the Communications Board within the chassis to meet