CN110456043A - 一种用于检测肺癌相关蛋白1的方法、试剂盒及其用途 - Google Patents
一种用于检测肺癌相关蛋白1的方法、试剂盒及其用途 Download PDFInfo
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- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
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Abstract
本发明涉及生物医药检测领域,具体涉及检测OLC1的方法、试剂盒及其用途。利用本发明提供的OLC1试剂盒和检测方法,可准确、灵敏、快速地测定样本中的OLC1浓度。
Description
技术领域
本发明涉及生物医药检测领域,具体涉及一种用于检测肺癌相关蛋白1的方法、试剂盒及其用途。
背景技术
肺癌相关蛋白1(overexpressed in lung cancer,OLC1)是一种新的肿瘤相关基因编码的蛋白,全长334个氨基酸,其Genbank ID:9798。通过生物信息学分析发现,肺癌相关蛋白OLC1可以在报告基因水平激活NF-kB通路。在肺鳞癌发生的早期阶段(不典型增生和原位癌),OLC1的表达量升高,提示其可能与肺癌的早期癌变关系密切。最近有研究表明,OLC1在多种人类恶性肿瘤,如卵巢癌、结直肠癌、食管癌、肺癌和乳腺癌中均高表达。
肺癌(lung cancer,LC)是中国乃至全世界最常见的恶性肿瘤以及首位的癌症死亡原因。据全国肿瘤登记中心的最新数据统计,2013年全国估计肺癌新发病例约73.28万例,占全部恶性肿瘤新发病例的19.90%,位居恶性肿瘤发病第一位。男性肺癌发病率普遍高于女性。目前市场上几种肺癌肿瘤标志物均表现为灵敏度和特异性较差。
乳腺癌(Breast cancer,BC),目前是中国女性发病率最高的癌症,癌症死亡原因位居第六。乳腺癌的早期发现和早期诊断是提高疗效和减少死亡的关键。虽然多种肿瘤标志物应用与乳腺癌的早期诊断,但有研究指出CEA、AFP、CA125、CA153、CA199这五中血清肿瘤标志物在乳腺癌早期诊断中的应用价值不高,采用单个血清肿瘤标志物来筛查乳腺癌,其敏感度均未超过10%,即使这五种标志物联合筛查,其敏感度也仅为20%。
鼻咽癌(nasopharyngeal carcinoma,NC)指发生于鼻咽腔顶部和侧壁的恶性肿瘤。是我国高发恶性肿瘤之一。鼻咽癌肿瘤标志物的应用和研究尚存在种种问题和困难。
目前,市场上尚未有检测OLC1相关的试剂盒,使用传统方法进行检测缺乏重复性、准确性,且纯手工加样等系列操作效率低,容易导致实验结果误差大,还易受外部干扰因素影响,不能形成产业化。
发明内容
本发明的目的在于提供一种检测肺癌相关蛋白1的方法、试剂盒及其用途。
本发明是通过以下技术方案实现的:
一种用于检测肺癌相关蛋白1的试剂盒,所述试剂盒为酶联免疫吸附试剂盒、化学发光试剂盒或POCT试剂盒;所述酶联免疫吸附试剂盒主要包括组分A1和组分B1,其中组分A1为吸附有第一抗OLC1抗体的微孔板,组分B1为标记有示踪标记物的第二抗OLC1抗体;所述化学发光试剂盒主要包括组分A2和组分B2,其中组分A2为结合于磁性微球上的第一抗OLC1抗体,组分B2为标记有示踪标记物的第二抗OLC1抗体;所述POCT试剂盒主要包括组分A3和组分B3,其中组分A3为结合于荧光微球上的第一抗OLC1抗体,组分B3为结合于固相载体的第二抗OLC1抗体;所述第一抗OLC1抗体和第二抗OLC1抗体与OLC1的结合位点不同。
优选地,所述组分A1所用的微孔板为塑料制品。
优选地,所述组分A1吸附第一抗OLC1抗体浓度为2-10μg/ml。
优选地,所述组分B1和B2的示踪标记物为碱性磷酸酶、辣根过氧化物酶、葡萄糖氧化酶中的至少一种。
优选地,所述组分B1和B2标记示踪物的方法为过碘酸钠法、戊二醛法中至少一种。
优选地,所述组分B1和B2标记示踪物的浓度为0.1-2mg/ml,所用第二抗OLC1抗体浓度为2-10μg/ml。
优选地,所述组分A2磁性微球为Fe2O3或Fe3O4磁性纳米粒子与有机高分子材料的复合体,粒径为0.1-2μm,并且微球表面经过活化而带有一种或多种活性功能基团。
优选地,所述组分A2所用第一抗OLC1抗体浓度为0.01-1mg/ml,所用磁性微球浓度为0.1-10mg/ml。
优选地,所述组分A3荧光微球为聚苯乙烯、聚氯乙烯纳米粒子与有机高分子材料的复合体,粒径为0.05-1μm,并且微球表面经过活化而带有一种或多种活性功能基团。
优选地,所述组分A3所用第一抗OLC1抗体浓度为0.01-1mg/ml,所用荧光微球浓度为0.1-5mg/ml。
优选地,所述组分B3所用的固相载体为孔径为1-10μm的硝酸纤维素膜。
优选地,所述标记有示踪标记物或结合于固相载体上的方式可以为直接或间接。
优选地,所述组分A2和A3结合微球的方式为共价结合。
优选地,所述试剂盒还包括肺癌相关蛋白1的低点校准品、高点校准品、空白对照品、质控品、标准液。
本发明还提供了一种检测肺癌相关蛋白1的方法,所述方法包括使用如上所述的试剂盒,通过免疫学原理检测待测样品中的肺癌相关蛋白1浓度。
优选地,使用酶联免疫吸附试剂盒包括以下步骤:将OLC1标准液、对照品、质控品和待测样本加入试剂盒组分A1中,经孵育、洗涤后加入组分B1,经孵育、洗涤后加入底物液,再加入终止液,检测吸光度,获得标准液与吸光度值之间的标准曲线,将检测得到的待测样本的吸光度值与标准曲线对照,计算得待测样本OLC1蛋白含量;或者
使用化学发光试剂盒包括以下步骤:将组分A2、B2与待测样本混匀,经孵育、洗涤、磁分离后加入发光底物液,检测光信号强度,获得标准液与光信号强度值之间的标准曲线,将检测得到的待测样本的光信号强度值与标准曲线对照,计算得待测样本OLC1蛋白含量;或者
使用POCT试剂盒包括以下步骤:将组分A3喷涂于结合垫上,将组分B3和组分A3一起组成试剂条,将样本直接加入到试剂条上,经孵育后直接检测光信号强度,获得标准液与光信号强度值之间的标准曲线,将检测得到的待测样本的光信号强度值与标准曲线对照,计算得待测样本OLC1蛋白含量。
本发明还提供了检测肺癌相关蛋白1在辅助诊断肺癌、乳腺癌和鼻咽癌中的用途。
本发明的有益效果在于:
根据本发明提供的OLC1检测试剂盒,包含以多种标记方式标记示踪标记物或者以多种方式结合于固相载体上的OLC1或OLC1抗体,利用双抗体夹心法或竞争法,可准确、灵敏、快速地测定样本中的OLC1浓度。
使用本发明提供的试剂盒测定OLC1,不需要将待测样本进行稀释处理,可以直接用于检测,从而有利于提高低值样本的检测灵敏度。其中待测样本可为空白管血清、分离胶管血清、促凝剂管血清、EDTA血浆和肝素血浆中的任意一种。
本发明提供的OLC1检测方法,可以采用酶联免疫、化学发光免疫、荧光免疫分析法等多种免疫学检测方法,试剂盒的特异性及灵敏度得到了很大的提升,更好的服务于临床诊断。
本发明提供的OLC1检测方法,使用所提供的试剂盒,借助免疫分析仪,加样方式完全由仪器全自动操作,降低了人为因素对实验结果造成的干扰,大幅度缩短了测试时间,有利于临床上快速的得到诊断结果。
本发明提供的检测肺癌相关蛋白1在辅助诊断肺癌、乳腺癌和鼻咽癌中的用途为癌症,特别是肺癌、乳腺癌和鼻咽癌的诊断提供了一个新的检测标志物,有利于癌症的早期发现和治疗。
附图说明
图1为本发明重组OLC1的SDS-PAGE检测结果图:其中,泳道M:蛋白Marker;泳道1:重组OLC1蛋白。
图2为本发明第一抗OLC1抗体和第二抗OLC1抗体的SDS-PAGE检测结果图,其中,泳道M:蛋白Marker;泳道1:第一抗OLC1抗体;泳道2:第二抗OLC1抗体。
图3为POCT试剂条简单构造图。
图4为OLC1区分正常人与肺癌患者(LC)的ROC曲线。
图5为OLC1区分正常人与乳腺癌(BC)的ROC曲线。
图6为OLC1区分正常人与鼻咽癌(NC)的ROC曲线。
具体实施方式
为更好理解本发明,下面结合实施例及附图对本发明作进一步描述,以下实施例仅是对本发明进行说明而非对其加以限定。
实施例1 OLC1重组蛋白的制备
1.构建重组菌:将OLC1的编码基因连接入基因的表达载体pGEX-4T-1,并将构建好的基因表达载体导入宿主大肠杆菌E.coli JM109(DE3)菌体中,构建重组菌JM109(DE3)/rOLC1;其中,编码OLC1的核苷酸序列如SEQ ID NO:1所示,OLC1的氨基酸序列如SEQ ID NO:2所示。
2.OLC1蛋白溶液的制备
(2-1)将重组OLC1工程菌接种到含有氨苄青霉素的LB固体培养基上,37℃恒温培养箱培养14~16h,挑取单菌落接种于3~5ml的LB培养基中,37℃恒温摇床培养至OD600达到1.5~1.8,转速为220r/min;
(2-2)将(2-1)获得的菌液1:100接种到100~200ml的LB培养基中,37℃恒温摇床培养OD600达到1.5~1.8,转速为220r/min,得到发酵种子液;
(2-3)将发酵种子液1:100接种于装有500ml已灭菌的LB培养基摇瓶中,设置温度为37℃、转速为220r/min条件下发酵;当摇瓶中液体的OD600达到1.0~1.2时,加入异丙基-β-D-硫代吡喃半乳糖苷至终浓度为1mmol/L,调温度至32℃,诱导表达4~5h,得到发酵液;
(2-4)将发酵液4℃条件下7000r/min离心15min,收集菌体,用适量的0.05M三羟甲基氨基甲烷-盐酸缓冲液重悬菌体,在功率为600~800W条件下超声破碎,超声时间1s,间隙时间1s,超声时间30min,4℃条件下12000r/min离心15min,收集上清,即可得到重组OLC1蛋白粗品;
(2-5)将OLC1蛋白粗品经过层析柱GSTrapTM4B(GE)亲和层析纯化获得纯化后OLC1蛋白;
(2-6)纯化后OLC1蛋白经过透析、过滤除菌即可得到OLC1蛋白。
3.OCL1蛋白SDS-PAGE鉴定,相对分子质量66kD(图1)。
实施例2 OLC1抗体的制备
采用大肠杆菌原核表达OLC1蛋白。取6-8周龄BALB/c小鼠,利用OLC1重组蛋白作为抗原免疫小鼠,免疫完成后再进行细胞融合、杂交瘤筛选、细胞克隆、用ELISA和Westernblot方法检测各细胞株分泌的抗体。经过鉴定后获得特异性和灵敏度最高的两株OLC1单克隆抗体杂交瘤细胞株,分别为分泌第一抗OLC1抗体(OLC1-38)和第二抗OLC1抗体(OLC1-15)的两株细胞株(图2),将这两株细胞直接免疫小鼠腹腔获得小鼠腹水,收货腹水后纯化抗体,即获得两株OLC1单克隆抗体。OLC1-38、OLC1-15(其保藏编号分别为15588和15587)两株杂交瘤细胞株保藏于中国科学医学院肿瘤医院。
实施例3制备OLC1检测的ELISA试剂盒
1.制备偶联辣根过氧化物酶的第二抗OLC1抗体,用经过优化的过碘酸钠法将辣根过氧化物酶通过共价键偶联于抗体上。
2.建立检测OLC1抗原的酶标双抗体夹心法。将第一抗OLC1抗体系列稀释包被96孔酶标板,100μl/孔,4℃过夜,洗板,用10%小牛血清做为封闭液,200μl/孔,37℃封闭1.5小时,洗板,将OLC1重组蛋白系列稀释后加入到酶标反应孔中,100μl/孔,37℃反应1.0小时,洗板,加入系列稀释的标有辣根过氧化物酶的第二抗OLC1抗体,100μl/孔,37℃反应1.0小时,洗板,加入TMB底物液,100μl/孔,37℃反应15-20min,加入2M硫酸终止反应,50μl/孔,用酶标仪OD450nm处检测吸光度;根据棋盘滴定结果确定OLC1的ELISA检测试剂盒的反应体系,再用该体系制备一批OLC1的ELISA检测试剂盒,检测待测样本,最终利用标准蛋白吸光度的值绘制标准曲线,并计算待测样本中OLC1的蛋白浓度。
实施例4制备OLC1检测的化学发光免疫试剂盒
1.制备偶联磁性微球的第一抗OLC1抗体,其中微球粒径0.5~3μm,包被浓度20-120μg抗体/1mg磁性微球,室温包被2h;磁分离包被完的磁性微球并用0.05M的BBST0.5%BSA封闭液封闭磁性微球30min;0.05M的BBST洗涤3次封闭完的磁性微球;0.01M PBS缓冲液重悬磁性微球待用。
2.制备偶联辣根过氧化物酶的第二抗OLC1抗体,用经过优化的过碘酸钠法将辣根过氧化物酶通过共价键偶联于抗体上。
3.试剂盒检测操作步骤:将偶联磁性微球的第一抗OLC1抗体、待测样本或OLC1标准蛋白、偶联辣根过氧化物酶的第二抗OLC1抗体同时加入反应杯,37°孵育10min,通过磁铁分离出夹心偶合物,磷酸盐缓冲液洗涤3次,加入发光底物,用光电倍增管检测发光值;利用标准蛋白的发光值及浓度绘制标准曲线,并计算待测样本中OLC1的含量。
实施例5制备OLC1检测的荧光免疫试剂盒
1.制备偶联荧光微球的第一抗OLC1抗体,微球粒径80~200nm,标记浓度10-120μg抗体/1mg荧光微球,室温或37°包被2h;离心分离包被完的磁性微球并用0.2M的BBST洗涤3次,0.5%BSA封闭液封闭磁性微球30min;0.05M的BBST洗涤3次封闭完的磁性微球;0.01MPBS缓冲液重选磁性微球待用。
2.通过喷金划膜仪器将偶联荧光微球的第一抗OLC1抗体喷涂于玻璃纤维制成的结合垫上,喷涂完毕烘烤干燥;将第二抗OLC1抗体和羊抗鼠抗体按照一定间距分别喷涂于硝酸纤维薄膜上,喷涂完毕干燥形成两条抗体线,喷涂第二抗OLC1抗体的抗体线作为T线(Test线),喷涂羊抗鼠抗体的抗体线作为C线(control线);将玻璃纤维制成的样品垫、喷涂了乳胶微球的结合垫、硝酸纤维薄膜、吸水纸按照图3所示依次粘贴于带粘胶的聚氯乙烯(PVC)背板上,形成检测试剂条,再将试剂条装入卡壳内。
3.荧光免疫层析实验操作步骤。将待测样本或标准蛋白由卡条样本孔滴入,50-120μl/条;室温孵育层析15min,然后将卡条插入荧光层析检测仪器中读取C\T线荧光值并计算T线与C线荧光值的比值;通过标准蛋白的T线和C线荧光比值及对应浓度绘制标准曲线,将样本的T\C线比值带入曲线获得待测样本中OLC1蛋白含量。
实施例6利用制备的试剂盒检测正常人与各组肿瘤患者血清样本中OLC1蛋白含量
利用制备好的OLC1化学发光试剂盒检测临床血清样本中OLC1蛋白含量,检测的样本包括731例正常人、842例肺癌患者、乳腺癌患者、鼻咽癌患者血清样本,检测结果显示肺癌、乳腺癌、鼻咽癌样本的OLC1蛋白浓度值显著高于正常人对照组(P<0.001,表1),检测数据如表1所示,对数据进行ROC曲线分析,当OLC1血清蛋白浓度在1.57ng/ml时,检测肺癌(Lung Cancer,LC)的敏感性为66.4%,特异性为80.3%,ROC曲线下面积为0.832(图4);当OLC1血清蛋白浓度在1.57ng/ml时,检测乳腺癌(Breast Cancer,BC)的敏感性为82.7%,特异性为80.3%,ROC曲线下面积为0.857(图5);当OLC1血清蛋白浓度在1.57ng/ml时,检测鼻咽癌(Nasopharynx Cancer,NC)的敏感性为74.3%,特异性为80.3%,ROC曲线下面积为0.843(图6);临床上可辅助诊断肺癌、乳腺癌和鼻咽癌。
表1 OLC1在正常人群和各组肿瘤患者血清中的浓度(ng/mL)
分组 | 例数 | 均值 | 中值 | 最大值 | 最小值 | 标准差 | P |
正常对照 | 583 | 1.63 | 0.78 | 80.9 | 0.02 | 4.77 | |
肺癌 | 842 | 4.49 | 1.8 | 183.53 | 0.12 | 15.69 | <0.001 |
乳腺癌 | 440 | 6.78 | 1.9 | 192.66 | 0.61 | 22.44 | <0.001 |
鼻咽癌 | 311 | 5.91 | 1.8 | 212.22 | 0.76 | 19.78 | <0.001 |
同样地,利用实施例3、4制备的ELISA试剂盒、荧光免疫试剂盒检测临床血清样本中OLC1蛋白含量,检测样本同上。结果显示所述试剂盒对检测肺癌、乳腺癌、以及鼻咽癌具有高的敏感性和特异性,临床上也可以用于辅助诊断肺癌、乳腺癌、以及鼻咽癌。
以上所述实施方式仅仅是对本发明的优选实施方式进行描述,并非对本发明的范围进行限定,在不脱离本发明设计精神的前提下,本领域普通技术人员对本发明的技术方案作出的各种变形和改进,均应落入本发明的权利要求书确定的保护范围内。
序列表
<110>中国医学科学院肿瘤医院;必欧瀚生物技术(合肥)有限公司
<120>一种用于检测肺癌相关蛋白1的方法、试剂盒及其用途
<130> 2017
<160> 2
<170> PatentIn version 3.3
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<211> 1008
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cgggaagact acctcgtgga ggccatggag atcctggagc tgtactgtga cctgctgctg 240
gctcggtttg gccttatcca gtctatgaag gaactagatt ctggtctggc tgaatctgtg 300
tctacattga tctgggctgc tcctcgactc cagtcagaag tggctgagtt gaaaatagtt 360
gctgatcagc tctgtgccaa gtatagcaag gaatatggca agctatgtag gaccaaccag 420
attggaactg tgaatgacag gctaatgcac aagctgagtg tggaagcccc acccaaaatc 480
ctggtggaga gatacctgat tgaaattgca aagaattaca acgtacccta tgaacctgac 540
tctgtggtca tggcagaagc tcctcctggg gtagagacag atcttattga tgttggattc 600
acagatgatg tgaagaaagg aggccctgga agaggaggga gtggtggctt cacagcacca 660
gttggtggac ctgatggaac ggtgccaatg cccatgccca tgcccatgcc tatgccatct 720
gcaaatacgc ctttctcata tccactgcca aagggaccag tagatgacat taatgctgat 780
aagaatatct cttctgcaca gattgttggt cctggaccca agccagaagc ctctgcaaag 840
cttccttcca gacctgcaga taactatgac aactttgtcc taccagagtt gccatctgtg 900
ccagacacac taccaactgc atctgctggt gccagcacct cagcatctga agacattgac 960
tttgatgatc tttcccggag gtttgaagag ctgaaaaaga aaacatag 1008
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Ala Asn Thr Pro Phe Ser Tyr Pro Leu Pro Lys Gly Pro Val Asp Asp
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Ile Asn Ala Asp Lys Asn Ile Ser Ser Ala Gln Ile Val Gly Pro Gly
260 265 270
Pro Lys Pro Glu Ala Ser Ala Lys Leu Pro Ser Arg Pro Ala Asp Asn
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Tyr Asp Asn Phe Val Leu Pro Glu Leu Pro Ser Val Pro Asp Thr Leu
290 295 300
Pro Thr Ala Ser Ala Gly Ala Ser Thr Ser Ala Ser Glu Asp Ile Asp
305 310 315 320
Phe Asp Asp Leu Ser Arg Arg Phe Glu Glu Leu Lys Lys Lys
325 330
Claims (17)
1.一种用于检测肺癌相关蛋白1(OLC1)的试剂盒,其特征在于:所述试剂盒为酶联免疫吸附试剂盒、化学发光试剂盒或POCT试剂盒;所述酶联免疫吸附试剂盒主要包括组分A1和组分B1,其中组分A1为吸附有第一抗OLC1抗体的微孔板,组分B1为标记有示踪标记物的第二抗OLC1抗体;所述化学发光试剂盒主要包括组分A2和组分B2,其中组分A2为结合于磁性微球上的第一抗OLC1抗体,组分B2为标记有示踪标记物的第二抗OLC1抗体;所述POCT试剂盒主要包括组分A3和组分B3,其中组分A3为结合于荧光微球上的第一抗OLC1抗体,组分B3为结合于固相载体的第二抗OLC1抗体;所述第一抗OLC1抗体和第二抗OLC1抗体与OLC1的结合位点不同。
2.根据权利要求1所述的试剂盒,其特征在于:所述组分A1所用的微孔板为塑料制品。
3.根据权利要求1所述的试剂盒,其特征在于:所述组分A1吸附第一抗OLC1抗体浓度为2-10μg/ml。
4.根据权利要求1所述的试剂盒,其特征在于:所述组分B1和B2的示踪标记物为碱性磷酸酶、辣根过氧化物酶、葡萄糖氧化酶中的至少一种。
5.根据权利要求1所述的试剂盒,其特征在于:所述组分B1和B2标记示踪物的方法为过碘酸钠法、戊二醛法中至少一种。
6.根据权利要求1所述的试剂盒,其特征在于:所述组分B1和B2标记示踪物的浓度为0.1-2mg/ml,所用第二抗OLC1抗体浓度为2-10μg/ml。
7.根据权利要求1所述的试剂盒,其特征在于:所述组分A2磁性微球为Fe2O3或Fe3O4磁性纳米粒子与有机高分子材料的复合体,粒径为0.1-2μm,并且微球表面经过活化而带有一种或多种活性功能基团。
8.根据权利要求1所述的试剂盒,其特征在于:所述组分A2所用第一抗OLC1抗体浓度为0.01-1mg/ml,所用磁性微球浓度为0.1-10mg/ml。
9.根据权利要求1所述的试剂盒,其特征在于:所述组分A3荧光微球为聚苯乙烯、聚氯乙烯纳米粒子与有机高分子材料的复合体,粒径为0.05-1μm,并且微球表面经过活化而带有一种或多种活性功能基团。
10.根据权利要求1所述的试剂盒,其特征在于:所述组分A3所用第一抗OLC1抗体浓度为0.01-1mg/ml,所用荧光微球浓度为0.1-5mg/ml。
11.根据权利要求1所述的试剂盒,其特征在于:所述组分B3所用的固相载体为孔径为1-10μm的硝酸纤维素膜。
12.根据权利要求1所述的试剂盒,其特征在于:所述标记有示踪标记物或结合于固相载体上的方式可以为直接或间接。
13.根据权利要求1所述的试剂盒,其特征在于:所述组分A2和A3结合微球的方式为共价结合。
14.根据权利要求1所述的试剂盒,其特征在于:所述试剂盒还包括肺癌相关蛋白1的低点校准品、高点校准品、空白对照品、质控品、标准液。
15.检测肺癌相关蛋白1的方法,其特征在于,所述方法包括使用如权利要求1所述的试剂盒,通过免疫学原理检测待测样品中的肺癌相关蛋白1浓度。
16.根据权利要求15所述的方法,其特征在于,使用酶联免疫吸附试剂盒包括以下步骤:将OLC1标准液、对照品、质控品和待测样本加入试剂盒组分A1中,经孵育、洗涤后加入组分B1,经孵育、洗涤后加入底物液,再加入终止液,检测吸光度,获得标准液与吸光度值之间的标准曲线,将检测得到的待测样本的吸光度值与标准曲线对照,计算得待测样本OLC1蛋白含量;或者
使用化学发光试剂盒包括以下步骤:将组分A2、B2与待测样本混匀,经孵育、洗涤、磁分离后加入发光底物液,检测光信号强度,获得标准液与光信号强度值之间的标准曲线,将检测得到的待测样本的光信号强度值与标准曲线对照,计算得待测样本OLC1蛋白含量;或者
使用POCT试剂盒包括以下步骤:将组分A3喷涂于结合垫上,将组分B3和组分A3一起组成试剂条,将样本直接加入到试剂条上,经孵育后直接检测光信号强度,获得标准液与光信号强度值之间的标准曲线,将检测得到的待测样本的光信号强度值与标准曲线对照,计算得待测样本OLC1蛋白含量。
17.检测肺癌相关蛋白1在辅助诊断肺癌、乳腺癌和鼻咽癌中的用途。
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Application publication date: 20191115 |
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