CN110702904A - 特定凝集素在制备用以鉴别肺癌分期的测试工具方面的用途以及装置 - Google Patents
特定凝集素在制备用以鉴别肺癌分期的测试工具方面的用途以及装置 Download PDFInfo
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- CN110702904A CN110702904A CN201910876927.4A CN201910876927A CN110702904A CN 110702904 A CN110702904 A CN 110702904A CN 201910876927 A CN201910876927 A CN 201910876927A CN 110702904 A CN110702904 A CN 110702904A
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Abstract
本发明提出了一种特定凝集素在制备用以鉴别肺癌分期的测试工具方面的用途以及装置。针对取自支气管肺泡灌洗液的样本,所述特定凝集素可分为三组:a组有RCA120、MAL‑II、EEL、PHA‑E、DBA、AAL;b组有PTL‑II、LCA、SJA、WFA、WGA;c组有PWM、VVA、GSL‑I。通过所述特定凝集素能够快速检测出差异性表达的糖蛋白糖链结构,确定受试者属于肺部良性病变、早期肺癌或晚期肺癌。本发明采用支气管肺泡灌洗液作为待检测物,由于其是从原发病灶、临近肿瘤细胞直接获取的样本,其具有高器官特异性,能直接反映患者身体的真实状况,且安全微创。
Description
技术领域
本发明涉及一种基于糖蛋白糖型鉴别肺癌分期的技术,具体涉及特定凝集素在制作这方面测试工具的用途以及相应的装置。
背景技术
肺癌是最常见的恶性肿瘤之一,严重危害人类健康,其发病率和致死性均居各恶性肿瘤榜首。《2018年全球癌症统计数据》和《2014年中国癌症的发病率和死亡率》报告显示,肺癌的发病率和死亡率占总癌症发病人口的11.6%和18.4%(全球)、20%和27.3%(中国)。
肺癌通常可分为非小细胞肺癌(Non-small cell lung cancer,NSCLC)和小细胞肺癌 (Small cell lung cancer,SCLC)。而根据病理学特征不同,NSCLC主要包括肺腺癌(Adenocarcinoma,ADC)和肺鳞癌(Squamous carcinoma,SCC)。肺癌早期临床症状的隐匿性,导致多数(57%)患者就诊时疾病已进展至晚期。由于较晚的诊断,肺癌的整体生存率较低,据统计,我国肺癌整体5年生存率仅为16.1%。目前应用于临床肺癌诊断的方法主要有影像学、细胞组织病理学及血清学指标检查,但因其不同程度的存在检出率低、高假阳性率、取材难度大或诊断效能不高等问题,临床实际应用效果欠理想,尤其早期诊断价值有限。支气管肺泡灌洗技术(BAL)是近几年在纤维支气管镜应用基础上发展起来的,为呼吸系统病变的诊断和疗效观察开辟了一条新途径。用于呼吸系统疾病诊断的支气管肺泡灌洗液(BALF)标本通过肺段、亚肺段灌洗术取得,可获得纤维支气管镜所不能探及的细胞学标本,提高癌细胞的检出率。肿瘤标志物是肿瘤细胞在发生、发展过程种中分泌、产生的一些化学活性物质,而且可通过肿瘤细胞分泌、释放于体液中,其实质反映了肿瘤的自身存在。研究认为,BALF中肿瘤标志物出现早、浓度高,是因为肺癌细胞分泌及代谢物首先进入支气管肺泡,后进入血循坏的缘故,且是从原发病灶获取的标本。目前已对肺癌患者BALF中的临床传统肿瘤标志物进行研究,认为其检测敏感性及特异性均高于血清学检测并在临床上广泛应用。
蛋白质的糖基化修饰广泛地存在于各种生物体中,是一种重要的翻译后修饰作用。有研究发现,异常糖基化的形成是肿瘤细胞恶性转变的一个关键特征,其反映了细胞表观遗传和参与多糖生物合成的基因异常表达。与健康组织相比,病变组织有例如很高程度的N- 糖链的分支型的变化、异构的岩藻糖基化、唾液酸化等糖基化变化,它们都可以作为某种疾病发生的生物指标。肿瘤糖基化的异常表达不是随机的,而是以常见的糖基化模式出现,其中甲胎蛋白(AFP)的岩藻糖基化的改变,对于检测高风险肝癌具有很高的敏感度。因此,将相关的糖蛋白用于精确诊断一些潜在的可能发展成为癌症的疾病,可被推广应用于发现糖链相关的生物标志物。在肺癌中己经发现很多异常糖基化,包括表达的改变、黏蛋白的糖基化、N-糖链分支类型改变、蛋白或糖脂上唾液酸化的增加等,但这些研究多集中在肺癌患者的组织或血清中。如何在保证特异性和灵敏度的前提下,尽量使用创伤低、易于获取的样本,拓展更多样本类型的研发工作是非常重要的。
凝集素芯片作为一种微阵列技术,凭借高通量、可重复性高、检测微量化、高度敏感及适用性广泛等优点,成为研究糖蛋白糖链结构变化最有效的分析工具之一,能够直接通过微量的原始样本快速、高灵敏地分析鉴定糖链,反映出样本中糖蛋白糖链最真实的状况。
发明内容
本发明通过大量的实验和分析,最终确立了针对支气管肺泡灌洗液作为待检测物,特定凝集素(组合)能够用来确定受试者肺癌分期情况。具体来说:
a组凝集素(可选以下凝集素任一或任意组合):RCA120、MAL-II、EEL、PHA-E、 DBA、AAL;
当凝集素RCA120、MAL-II、EEL、PHA-E所识别的糖链结构显著上调,凝集素DBA、AAL所识别的糖链结构显著下调,则判定样本来自肺部良性病变患者;反之,则判定样本来自肺癌早期或肺癌晚期患者。
b组凝集素(可选以下凝集素任一或任意组合):PTL-II、LCA、SJA、WFA、WGA;
当凝集素PTL-II、LCA、SJA、WFA所识别的糖链结构显著下调,凝集素WGA所识别的糖链结构显著上调,则判定样本来自肺癌早期患者;反之,则判定样本来自肺部良性病变患者或者肺癌晚期患者。
c组凝集素(可选以下凝集素任一或任意组合):PWM、VVA、GSL-I;
当凝集素PWM所识别的糖链结构显著下调,凝集素VVA、GSL-I所识别的糖链结构显著上调,则判定样本来自肺癌晚期患者;反之,则判定样本来自肺部良性病变患者。
还可设置d组凝集素,用于辅助验证a、b、c组凝集素的测试结果;d组凝集素为以下凝集素任一或任意组合:MAL-II、RCA120、ECA、HHL、GSL-I、DBA、DSA、AAL;
当凝集素MAL-II、RCA120、ECA、HHL所识别的糖链结构显著上调,凝集素GSL-I、DBA、DSA、AAL所识别的糖链结构显著下调,则判定样本来自肺部良性病变(BPD)患者;反之,则判定样本来自肺癌患者。
测试工具(载体)优选凝集素芯片,也可以是酶标板、磁粒等。
以凝集素芯片的固定为例:凝集素芯片是将各种不同来源的凝集素固定于醛基化、环氧化或经其他方式修饰后的玻璃片基上,再与标记后的糖蛋白、菌体、细胞等待检测样品反应,以检测待检测样品的糖链结构。
以环氧化磁粒为例:合成Fe3O4磁性微粒,然后用硅烷化试剂修饰磁性微粒制备得到环氧化磁性微粒。合成的环氧化磁性微粒被应用于凝集素等蛋白的固定化。
本发明通过特定凝集素能够快速检测出差异性表达的糖蛋白糖链结构,确定受试者属于肺部良性病变、早期肺癌或晚期肺癌。
本发明采用支气管肺泡灌洗液作为待检测物,与传统的血液样本相比,由于其是从原发病灶、临近肿瘤细胞直接获取的样本,其具有高器官特异性,能直接反映患者身体的真实状况,且安全微创,可广泛应用于临床肺部相关疾病的筛查及诊断。
基于本发明,可针对性地选取凝集素制作凝集素芯片等测试工具,一定程度上也降低了测试成本。
附图说明
图1为凝集素芯片点样阵列图,实验所用芯片包含37种凝集素,2个阴性质控BSA,1个阳性质控Marker。每张芯片上有四个重复区域,每个区域的每种凝集素重复3次,即每个区域由12*10的微阵列构成。
图2为各患者支气管肺泡灌洗液凝集素芯片荧光检测结果,其中,各图依次对应于肺部良性病变患者(BPD)、肺腺癌患者(ADC)、肺鳞癌(SCC)、小细胞肺癌患者(SCLC)、肺部良性病变患者(BPD)、肺癌早期患者(LC-ES)、肺癌晚期患者(LC-AS)。
图3为各患者(BPD、ADC、SCC、SCLC)支气管肺泡灌洗液中差异凝集素个例芯片荧光信号归一化数据图。
图4为各患者(BPD、LC-ES、LC-AS)支气管肺泡灌洗液中差异凝集素个例芯片荧光信号归一化数据图。
图5为BPD、LC-ES、LC-AS三组支气管肺泡灌洗液混合样本凝集素印迹验证结果图。其中(a)为SDS-PAGE银染结果图,(b)为凝集素RCA120的lectin blotting图,(c) 为具有明显差异的蛋白质条带灰度值分析图。
具体实施方式
以下介绍了本申请相关验证实验及分析,发明人具体的研发过程不限于此;给出的具体实施例也并非对本申请的限定。
本申请从37种凝集素(如表1所示)中筛选用于鉴别肺癌患者的凝集素探针。
表1 37种凝集素名称
本申请中主要使用的试剂材料如表2所示,其余常用试剂均为国产分析纯级别;仪器设备如表3所示,其余常用仪器设备均为国产。
表2实验所用试剂与材料
表3实验所用仪器与设备
1.研究人群和支气管肺泡灌洗液采集
符合条件的肺癌患者不能同时罹患其他疾病,且均为初诊患者,即未采取任何治疗措施。医院专业人员在患者行纤维支气管镜检查时采集肺部良性病变患者(BPD)、肺腺癌患者(ADC)、肺鳞癌患者(SCC)、小细胞肺癌患者(SCLC)支气管肺泡灌洗液样本各数例(如表4所示)。总共有281例样本,在进行分期研究时,去除了未分期的46例样本。
样本采集过程为先行术前准备,然后所有患者均予以纤维支气管镜经鼻插入,先检查左右支气管,并根据患者胸部CT显示的病变部位,将纤维支气管镜远端楔于病变所在的段支气管开口处,弥漫病变则冲洗右中叶或左舌叶,注入37℃温生理盐水,10-15ml/次,灌洗2次,以13kPa负压尽可能吸尽灌洗液,收集约10-15ml肺泡灌洗液于无菌管中。
表4样本信息汇总表
注:BPD,肺部良性病变;ADC,肺腺癌;SCC,肺鳞癌;SCLC,小细胞肺癌;X,未分期;LD,局限期;ED,广泛期。
2.支气管肺泡灌洗液蛋白处理和荧光标记
收集到的支气管肺泡灌洗液经4000rpm,4℃离心20min后收集上清液,再经AmiconUltra-4 3KD滤膜对上清液进行浓缩、除盐及纯化,以1ul/ml加入蛋白酶抑制剂,然后用Brandford法定量样本蛋白浓度,分装后保存于-80℃冰箱中或用干冰运输。各个例样本经Cy3荧光染料标记后用Sephadex G-25除盐柱去掉游离荧光,收集到的荧光标记后的支气管肺泡灌洗液蛋白用于凝集素芯片孵育。另外将所采集样本按照肺部良性病变组(BPD)、肺癌早期组(LC-ES)及肺癌晚期组(LC-AS)各取100μl进行混合,混合后的三组样本用于凝集素印迹实验。
3.凝集素芯片及其数据分析
凝集素芯片的制备,Cy3荧光标记的样本蛋白与凝集素芯片的孵育步骤及凝集素芯片数据获取与归一化分析与专利申请号为201110021447.3发明专利描述的凝集素芯片和数据分析过程一致。
4.凝集素印迹及其数据分析
采用Bradford法定量BPD、LC-ES、LC-AS三组支气管肺泡灌洗液混合样本中蛋白浓度,并取含5μg蛋白的样本同5×上样缓冲液混合,置于100℃沸水中水浴加热5min使其完全变性,而后立即冰上冷却、离心。配制3%的浓缩胶及10%聚丙烯酰胺分离胶进行蛋白质电泳。完成电泳后的凝胶采用银染方法进行染色,一方面对三组支气管肺泡灌洗液混合样本中的蛋白分子量及丰度分布有所了解,另一方面检测蛋白质浓度定量是否准确,证明用于后续凝集素印迹实验的蛋白上样量是否一致。另取25μg样本蛋白进行SDS-PAGE 电泳,后采用湿转仪将蛋白转至PVDF膜上,经过Carbo-free试剂封闭PVDF膜1h后,加入Cy5荧光标记的凝集素至终浓度2μg/mL,4℃避光震荡过夜。孵育结束后在避光条件下用TBST缓冲液清洗PVDF膜四次,每次10min,然后在Storm 840多功能图像分析仪上设置参数PMT为800,高分辨率,激发光波长635nm下扫描图像,图像利用ImageJ软件分析凝集素-蛋白条带灰度值。
5.凝集素芯片结果分析
(1)肺部良性病变患者、肺腺癌患者、肺鳞癌患者及小细胞肺癌患者间支气管肺泡灌洗液糖蛋白糖型的比较
利用凝集素芯片分别对肺部良性病变患者、肺腺癌患者、肺鳞癌患者及小细胞肺癌患者支气管肺泡灌洗液样本进行检测(见图2),通过专业软件获取芯片数据并归一化处理后,将四组个例样本凝集素芯片结果进行比较(见图3)。
结果发现,对于支气管肺泡灌洗液样本,肺部良性病变患者(BPD)与上述三种类型肺癌患者相比,有8种凝集素识别的糖链均存在差异性表达;而在上述三种类型肺癌患者之间无显著性差异。其中MAL-II识别的Siaα2-3Galβ1-4Glc(NAc)/Glc,Siaα2-3Gal,Siaα2-3,和Siaα2-3GalNAc糖链;RCA120识别的β-Gal,Galβ-1,4GlcNAc(type II)和Galβ1-3GlcNAc(type I)糖链;ECA识别的Galβ-1,4GlcNAc(type II)和Galβ1-3GlcNAc(type I)糖链;HHL识别的High-Mannose,Manα1-3Man,Manα1-6Man和Man5-GlcNAc2-Asn糖链结构在BPD 患者支气管肺泡灌洗液中表达量高于患有肺癌的支气管肺泡灌洗液中的表达量,相反, GSL-Ⅰ识别的αGalNAc,αGal,anti-A and B糖链;DBA识别的αGalNAc,Tn antigen和 GalNAcα1-3((Fucα1-2))Gal(blood group A antigen)糖链;AAL识别的Fucα1-6GlcNAc(corefucose),和Fucα1-3(Galβ1-4)GlcNAc糖链;DSA识别的β-D-GlcNA,(GlcNAcβ1-4)n, Galβ1-4GlcNAc糖链结构在不同类型肺癌的支气管肺泡灌洗液中表达量均升高。
有5种凝集素识别的糖链在ADC中同BPD及SCC患者支气管肺泡灌洗液相比均存在差异性表达,而与SCLC患者支气管肺泡灌洗液之间无显著性差异。其中PHA-E识别的Bisecting GlcNAc,biantennary complex-type N-glycan with outer Gal糖链;EEL识别的Galα1-3(Fucα1-2)Gal(blood group B antigen)糖链;BPL识别的Galβ1-3GalNAc,Terminal GalNAc糖链结构在ADC患者支气管肺泡灌洗液中的表达量高于BPD及SCC患者;而凝集素GSL-II和VVA分别识别的GlcNAc and agalactosylated tri/tetra antennaryglycans糖链和terminal GalNAc,GalNAcα-Ser/Thr(Tn),GalNAcα1-3Gal糖链结构在ADC中则高表达。
凝集素PNA识别的Galβ1-3GalNAcα-Ser/Thr(T)糖链在ADC患者的支气管肺泡灌洗液中较BPD及SCC患者高表达,而在SCC患者的支气管肺泡灌洗液中较ADC及SCLC患者则低表达。
另外,MPL凝集素识别的Galβ1-3GalNAc,GalNAc糖链结构在SCC患者支气管肺泡灌洗液中较BPD及SCLC患者低表达,而与ADC患者的支气管肺泡灌洗液表达情况无显著差异。
(2)肺部良性病变患者、肺癌早期患者、肺癌晚期患者支气管肺泡灌洗液糖蛋白糖型的比较
本申请进一步将所收样本按照肺部良性病变患者(BPD)、肺癌早期患者(LC-ES,非小细胞肺癌的Ⅰ/Ⅱ期和小细胞肺癌的局限期)、肺癌晚期患者(LC-AS,非小细胞肺癌的Ⅲ/Ⅳ期和小细胞肺癌的广泛期)进行分组,利用凝集素芯片对三组的个例样本支气管肺泡灌洗液样本进行检测(见图2),通过专业软件获取芯片数据并归一化处理后,将三组个例样本凝集素芯片结果进行比较(见图4)。
结果发现,有6种凝集素识别的糖链在BPD中同不同时期肺癌患者支气管肺泡灌洗液相比均存在差异性表达,而在不同时期肺癌患者支气管肺泡灌洗液间则无明显差异。其中 RCA120识别的β-Gal,Galβ-1,4GlcNAc(type II)和Galβ1-3GlcNAc(type I)糖链;MAL-II 识别的Siaα2-3Galβ1-4Glc(NAc)/Glc,Siaα2-3Gal,Siaα2-3,和Siaα2-3GalNAc糖链;EEL识别的FGalα1-3(Fucα1-2)Gal(blood group B antigen)糖链;PHA-E识别的BisectingGlcNAc, biantennary complex-type N-glycan with outer Gal糖链结构无论是在肺癌早期患者支气管肺泡灌洗液中,还是在肺癌晚期患者支气管肺泡灌洗液中,其表达量均低于BPD患者支气管肺泡灌洗液中的表达量;相反的是,DBA识别的αGalNAc,Tn antigen和
GalNAcα1-3((Fucα1-2))Gal(blood group A antigen)糖链;AAL识别的Fucα1-6GlcNAc(core fucose),和Fucα1-3(Galβ1-4)GlcNAc糖链结构在不同时期肺癌的支气管肺泡灌洗液中表达量均升高。
有4种凝集素识别的糖链在肺癌早期患者支气管肺泡灌洗液中表达量显著降低相较于 BPD及肺癌晚期组,分别是PTL-II识别的Gal,blood group H,T-antigen糖链;LCA识别的α-D-Man,Fucα-1,6GlcNAc,α-D-Glc糖链和SJA识别的αGalNAc,αGal,anti-A andBTerminal in GalNAc and Gal,anti-A and anti-B human blood group糖链;WFA识别的terminating in GalNAcα/β1-3/6Gal糖链结构。而WGA凝集素识别的Multivalent Sia and(GlcNAc)n糖链结构在肺癌早期患者支气管肺泡灌洗液中表达量显著升高。
凝集素PWM识别的Branched(LacNAc)n糖链在肺癌晚期患者支气管肺泡灌洗液中同 BPD及肺癌早期组相比为低表达,而凝集素VVA识别的terminal GalNAc, GalNAcα-Ser/Thr(Tn),GalNAcα1-3Gal糖链在肺癌晚期患者中则高表达。
另有1种凝集素GSL-I,其识别的αGalNAc,αGal,anti-A and B糖链结构随着患者病情的进展,表达量逐渐升高。
6.凝集素印迹验证结果
为进一步验证BPD、LC-ES、LC-AS三组支气管肺泡灌洗液凝集素芯片的结果,选择特异性的凝集素分别进行凝集素印迹实验,如图5,以凝集素RCA120为例,将Cy5荧光标记的RCA120同转膜后的三组支气管肺泡灌洗液蛋白孵育,经扫描仪扫描并测量其荧光条带灰度结果显示,RCA120与LC-ES组和LC-AS组支气管肺泡灌洗液中分子量在85kDa、 52-60kDa、45kDa的蛋白结合信号低于其与BPD组支气管肺泡灌洗液中对应分子量的蛋白质的结合信号,与凝集素芯片结果一致。
7.凝集素探针组的确定
通过上述结果分析,用于筛选不同类型及分期肺癌的凝集素探针被确定,↑为表达上调;↓为表达下调;—为无显著性差异。
(1)不同类型肺癌凝集素探针组的确定
若以肺部良性病变患者支气管肺泡灌洗液作为对照标准,区别于肺癌患者的凝集素探针表达结果参见表5。
表5用于将肺部良性病变患者区别于肺腺癌、肺鳞癌、小细胞肺癌患者的凝集素探针
由于通常对于肺部有病变的患者才有必要留取支气管肺泡灌洗液进行样本检测,所以此类测试自然预先排除了健康人群。因此,该组凝集素探针可以用来鉴别受试者是否患有肺癌(包括肺腺癌、肺鳞癌、小细胞肺癌患者)。
若以肺腺癌患者支气管肺泡灌洗液作为对照标准,区别于肺部良性病变及肺鳞癌患者支气管肺泡灌洗液的凝集素探针表达结果参见表6。
表6用于将肺腺癌患者区别于肺部良性病变及肺鳞癌患者的凝集素探针
若以肺鳞癌患者支气管肺泡灌洗液作为对照标准,区别于肺部良性病变、肺腺癌及小细胞患者支气管肺泡灌洗液的凝集素探针表达结果参见表7。
表7用于将肺鳞癌患者区别于肺部良性病变、肺腺癌及小细胞肺癌患者中的凝集素探针
(2)不同分期肺癌凝集素探针组的确定
若以肺部良性病变患者支气管肺泡灌洗液作为对照标准,区别于不同分期肺癌患者支气管肺泡灌洗液的凝集素探针表达结果参见表8。
表8用于将肺部良性病变患者区别于肺癌早期、肺癌晚期患者的凝集素探针
若以肺癌早期患者支气管肺泡灌洗液作为对照标准,区别于肺部良性病变及肺癌晚期患者支气管肺泡灌洗液的凝集素探针表达结果参见表9。
表9用于将肺癌早期患者区别于肺部良性病变及肺癌晚期患者的凝集素探针
若以肺癌晚期患者支气管肺泡灌洗液作为对照标准,区别于肺部良性病变及肺癌早期患者支气管肺泡灌洗液的凝集素探针表达结果参见表10。
表10用于将肺癌晚期患者区别于肺部良性病变及肺癌早期患者的凝集素探针
本申请采用支气管肺泡灌洗液作为待检测物,与传统的血液样本相比,由于其是从原发病灶、临近肿瘤细胞直接获取的样本,其具有高器官特异性,能直接反映患者身体的真实状况,且安全微创,可广泛应用于临床肺部相关疾病的筛查及诊断。基于本申请,可针对性地选取凝集素制作凝集素芯片等测试工具,一定程度上也降低了测试成本。
Claims (6)
1.特定凝集素在制备用以鉴别肺癌分期的测试工具方面的用途,其特征在于:针对取自支气管肺泡灌洗液的样本,所述特定凝集素为以下至少一组凝集素:
a组凝集素为以下凝集素任一或任意组合:RCA120、MAL-II、EEL、PHA-E、DBA、AAL;
当凝集素RCA120、MAL-II、EEL、PHA-E所识别的糖链结构显著上调,凝集素DBA、AAL所识别的糖链结构显著下调,则判定样本来自肺部良性病变患者;反之,则判定样本来自肺癌早期或肺癌晚期患者;
b组凝集素为以下凝集素任一或任意组合:PTL-II、LCA、SJA、WFA、WGA;
当凝集素PTL-II、LCA、SJA、WFA所识别的糖链结构显著下调,凝集素WGA所识别的糖链结构显著上调,则判定样本来自肺癌早期患者;反之,则判定样本来自肺部良性病变患者或者肺癌晚期患者;
c组凝集素为以下凝集素任一或任意组合:PWM、VVA、GSL-I;
当凝集素PWM所识别的糖链结构显著下调,凝集素VVA、GSL-I所识别的糖链结构显著上调,则判定样本来自肺癌晚期患者;反之,则判定样本来自肺部良性病变患者。
2.根据权利要求1所述的用途,其特征在于:所述特定凝集素还包括d组凝集素,用于辅助验证a、b、c组凝集素的测试结果;d组凝集素为以下凝集素任一或任意组合:MAL-II、RCA120、ECA、HHL、GSL-I、DBA、DSA、AAL;
当凝集素MAL-II、RCA120、ECA、HHL所识别的糖链结构显著上调,凝集素GSL-I、DBA、DSA、AAL所识别的糖链结构显著下调,则判定样本来自肺部良性病变(BPD)患者;反之,则判定样本来自肺癌患者。
3.根据权利要求1或2所述的用途,其特征在于:所述测试工具为凝集素芯片。
4.一种用于肺癌分期的筛查、早期诊断、风险评估、药物筛选和/或疗效评估的装置,针对取自支气管肺泡灌洗液的样本,其特征在于,包括:
A、用于获取特定糖蛋白糖链结构的表达水平的设备,所述特定糖蛋白糖链结构对应于权利要求1或2中所述的特定凝集素;
B、用于判别所述特定糖蛋白糖链结构是否显著上调/下调的标识、模块或处理器。
5.根据权利要求4所述的装置,其特征在于:所述用于获取特定糖蛋白糖链结构的表达水平的设备包括凝集素芯片、孵育盒和生物芯片扫描系统,其中凝集素芯片上设置有相应的所述特定凝集素。
6.根据权利要求4所述的装置,其特征在于:所述标识、模块或处理器预先记录有肺部良性病变(BPD)、肺腺癌(ADC)、肺鳞癌(SCC)、小细胞肺癌(SCLC)、肺癌早期(LC-ES)和肺癌晚期(LC-AS)对应的参考值,用于与样本结果对照确定是否显著上调/下调。
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