CN109313195A - 用于提供胆道癌诊断信息的方法和用于诊断胆道癌的装置 - Google Patents

用于提供胆道癌诊断信息的方法和用于诊断胆道癌的装置 Download PDF

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CN109313195A
CN109313195A CN201780036260.6A CN201780036260A CN109313195A CN 109313195 A CN109313195 A CN 109313195A CN 201780036260 A CN201780036260 A CN 201780036260A CN 109313195 A CN109313195 A CN 109313195A
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bile ducts
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俞炳掇
金庚喜
禹尚命
孔善映
金泰现
朴商在
李宇振
韩盛植
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National Cancer Center Korea
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Abstract

本发明涉及提供胆道癌诊断信息的方法和用于诊断胆道癌的装置。根据本发明的提供胆道癌诊断信息的方法包括以下步骤：获取生物样品；测量所述生物样品中胆道癌预测标记物的浓度；以及从测定的所述标记物的浓度提供胆道癌诊断信息，所述标记物包括Nudifloramide。

Description

用于提供胆道癌诊断信息的方法和用于诊断胆道癌的装置
技术领域
本发明涉及用于提供胆道癌诊断信息的方法和用于诊断胆道癌的装置。
背景技术
癌是通过细胞的无限增殖妨碍正常细胞功能的疾病，代表性的癌症实例包括肺癌、胃癌(GC)、乳腺癌(BRC)、结肠直肠癌(CRC)、胆道癌以及卵巢癌(OVC)等，但癌实质上可以在任何组织中发生。
已针对早期诊断癌症付出了很多的努力，对于胆道癌，有腹部CT、CA19-9电化学发光免疫分析(ECLIA)和甲胎蛋白检查等，但还未开发出准确度高且非侵袭式的诊断方法。
发明内容
技术课题
本发明的一方面涉及提供准确度高且非侵袭式的用于提供胆道癌诊断信息的方法和用于诊断胆道癌的装置。
课题解决手段
根据本发明的一方面，提供了一种用于提供胆道癌诊断信息的方法，包括：获取生物样品；测量所述生物样品中用于预测胆道癌的标记物的浓度；以及使用测量的所述标记物的浓度提供针对胆道癌的诊断信息。所述标记物包括Nudifloramide。
所述标记物还可以包括LPC18:0和纤维蛋白原α链中的至少一个。
用于胆道癌诊断的Nudifloramide的标准浓度是220pg/μl至320pg/μl。
所述纤维蛋白原α链的标准浓度是130pg/μl至200pg/μl。
所述LPC18:0的浓度可以利用质谱法测量，用于胆道癌诊断的所述LPC18:0的标准浓度是1,500,000至2,000,000au。
在提供信息的步骤中，确定是否是胆道癌可以基于Nudifloramide和纤维蛋白原α链的特定浓度或更高浓度，以及LPC18:0的特定浓度或者更低浓度。
所述生物样品可以包括血清。
所述浓度可以通过质谱法测量。
根据本发明的一方面，提供了一种用于诊断胆道癌的装置，包括：被构造为输入从生物样品中检测的质谱数据的输入单元；以及被构造为从所述质谱数据计算用于预测胆道癌的标记物的浓度并基于计算出的浓度确定针对胆道癌的诊断信息的诊断单元。所述标记物包括Nudifloramide。
所述标记物还可以包括LPC18:0和纤维蛋白原α链中的至少一个。
用于胆道癌诊断的Nudifloramide的标准浓度是220pg/μl至320pg/μl。
根据本发明的另一个方面，提供了一种用于胆道癌诊断的方法，包括：获取生物样品；测量所述生物样品中用于预测胆道癌的标记物的浓度；以及基于测量的所述标记物的浓度诊断胆道癌。所述标记物包括Nudifloramide。
所述标记物还可以包括LPC18:0和纤维蛋白原α链中的至少一个。
用于胆道癌诊断的Nudifloramide的标准浓度可以是220pg/μl至320pg/μl。
用于胆道癌诊断的所述纤维蛋白原α链的标准浓度可以是130pg/μl至200pg/μl。
LPC 18:0的浓度可以利用质谱仪测量，用于胆道癌诊断的LPC18:0的标准浓度是1,500,000au至2,000,000au。
在诊断胆道癌的步骤中，确定是否是胆道癌可以基于Nudifloramide和所述纤维蛋白原α链的特定浓度或者更高浓度，并基于LPC18:0的特定浓度或者更低浓度。
所述生物样品可以包括血清。
所述浓度通过质谱法测定。
发明效果
根据本发明的实施方案的用于诊断针对胆道癌的诊断信息的方法以及用于诊断胆道癌的装置可以具有高的准确度高且是非侵袭式的。
附图说明
图1示出根据本发明的一个实施方案的用于诊断胆道癌的装置。
图2示出利用nudifloramide浓度的诊断结果。
图3示出利用LPC18:0浓度的诊断结果。
图4示出利用LPC18:0浓度的另一个诊断结果。
图5示出利用纤维蛋白原α链浓度的诊断结果。
具体实施方式
本发明中，术语“生物样品”包括全血、血清、血浆、尿、粪便、痰、唾液、组织、细胞、细胞提取物、体外细胞培养物等样品，但不限于此。
本发明是以Nudifloramide可用作胆道癌的标记物的发现为依据。
Nudifloramide的正式名称是N-甲基-2-吡哆酮-5-甲酰胺(N-Met hyl-2-pyridoxone-5-carboxamide)；1,6-二氢-1-甲基-6-氧代烟酰胺(1,6-Dihydro-1-methyl-6-oxonicotinamide)；3-氨甲酰基-1-甲基-6-吡啶酮(3-Carbamoyl-1-methyl-6-pyridone)，结构式如下。
作为胆道癌的标记物，还可以利用溶血性磷脂酰氯(LPC,lysophospatidychloride)18:0和/或纤维蛋白原α链(FAC)。
测量生物样品例如血清中的标记物的浓度(浓度值)，并从测量的浓度可以诊断是否是胆道癌。浓度测量可以利用质谱仪进行，但并不限于此，利用质谱仪测定的标记物相应的任意单位(“au”)值测定每一标记物的浓度。
LPC18:0和纤维蛋白原α链的作用是提高诊断结果的特异性、阳性预测值(PPV)和/或阴性预测值(NPV)。
在胆道癌诊断中，对于Nudifloramide和FAC，如果Nudifloramide和FAC的浓度高于或者等于每个特定浓度，则可以确定为胆道癌，对于LPC18:0，如果LPC18:0的浓度小于或者等于特定浓度，则可以确定为胆道癌。
关于Nudifloramide，作为胆道癌诊断的标准的特定浓度可以是150pg/μl至500pg/μl，200pg/μl至350pg/μl，220pg/μl至320pg/μl或者250pg/μl至300pg/μl之间。
关于FAC，作为胆道癌诊断的标准的特定浓度可以在100pg/μl至250pg/μl，130pg/μl至200pg/μl或者150pg/μl至180pg/μl之间。
关于LPC18:0，作为胆道癌诊断的标准的特定浓度是质谱仪的au值，可以在1,000,000至2,500,000，1,500,000至2,000,000或者1,600,000至1,800,000之间。
同时，上面提示的特定浓度范围是根据本发明的实施例的条件下的值。如果一个具体浓度范围在使用其他采样条件、预处理条件或其他仪器时在上述范围外，但如果其遵从本公开的条件将落入特定浓度范围内，则应理解其属于本公开的权利范围内。
下面参考附图对本发明进行更详细地说明。
附图仅仅是为了更详细地说明本发明的技术思想图示的一例，因此，本发明的思想并不局限于附图。
图1是示出根据本发明的实施方案的用于诊断胆道癌的装置。
输入单元(100)中输入从生物样品检测出的质谱数据(以下称为“诊断对象数据”)。
诊断单元(200)从诊断对象数据测量标记物的浓度，并根据测量的浓度生成针对胆道癌的诊断信息。即，在诊断单元(200)中对诊断对象数据确定胆道癌阳性或胆道癌阴性。在此过程中，诊断单元(200)可以仅使用Nudifloramide浓度，或者进一步使用LPC18:0和纤维蛋白原α链的浓度中的至少一个。
诊断单元(200)中，对于各标记物设定了标准浓度，标准浓度根据诊断对象者的各种信息比如年龄、性别、是否具有其他癌等可进行变更。
输出单元(300)输出针对胆道癌的诊断信息，可以利用显示器。
下面通过实验例对本发明进行更详细地说明。
获得血清
从92名胆道癌患者、34名胰腺癌患者、100名肺癌患者、30名胃癌患者、3名卵巢癌患者、340名正常对照组获得血清。
血清的提取
利用改进的布莱-戴尔法(Bligh and dyer method)提取了胆道癌患者和其他种类癌症患者以及正常对照组的血清。具体地方法如下。
在50μl的血清中添加1ml的蒸馏水之后，添加2ml的甲醇和0.9ml的二氯甲烷。
完全混合后，在冰上放置30分钟，然后再添加1ml的蒸馏水和0.9ml的二氯甲烷。
然后，在常温下进行1,500rpm持续10分钟的离心，分离上层液后用氮气进行干燥。
定量分析(浓度测定)
把提取样品5μl注入到Nexera X2 LC系统(Shimadzu)，利用待分析溶剂的浓度梯度(溶剂A，0.1％FA水溶液；溶剂B，100％ACN；1％溶剂B持续1.5min，1％-25％溶剂B持续4.5min，25％-45％溶剂B持续2min，45％-90％溶剂B持续2min，90％溶剂B持续4min，90％-1％溶剂B持续0.5min并在1％溶剂B中持续5.5min)分离后，使用阳离子MRM模式的TripleTOF 5600+系统(SCIEX)质谱仪进行了定量分析。
Nudifloramide
各种癌和正常对照群中的Nudifloramide的浓度分布如图2，以270pg/μl为标准确定的结果如表1。
[表1]
基于表1中的确定结果，基于270pg/μl Nudifloramide的胆道癌的灵敏度、特异性、PPV和NPV如下。
灵敏度：80.43％(74/92)
特异性：91.71％((28+310+98+28+56)/(34+340+100+30+63))
阳性结果值(PPV)：61.16％(74/(74+6+30+2+2+7))
阴性结果值(NPV)：96.65％
((28+310+98+28+56)/(18+28+310+98+28+56))
从上面的结果看出，单独使用Nudifloramide就能实现高的灵敏度、特异性、PPV及NPV。
LPC 18:0
各种癌和正常对照组中的LPC18:0的浓度分布如图3，基于质谱仪au1,000,000的确定结果如表2。
[表2]
基于表2中的确定结果，LPC18:0的基于质谱仪au1,000,000的胰腺癌的灵敏度、特异性、PPV和NPV如下。
灵敏度：97.06％(33/34)
特异性：82.08％((39+329+92+17+36)/(92+340+100+30+63))
阳性预测值(PPV)：22.76％(33/(53+33+11+8+13+27))
阴性预测值(NPV)：99.81％(1/(39+1+329+92+17+36))
把LPC18:0的标准变更为质谱仪au1,700,000时的结果如图4及表3。
[表3]
基于表3中的确定结果，基于LPC18:0的质谱仪au1,700,000的胰腺癌的灵敏度、特异性、PPV和NPV如下。
灵敏度：100％(34/34)
特异性：61.44％((18+266+64+11+25)/(92+340+100+30+63))
阳性预测值(PPV)：12.36％(34/(74+34+74+36+19+38))
阴性预测值(NPV)：100％
((18+266+64+11+25)/(18+0+266+64+11+25))
从上面的结果可以知道，LPC18:0是对胰腺癌有用的标记物。另一方面，LPC 18:0的标准浓度高时，不仅是胰腺癌、而且是胆道癌的标记物，如下文所述。
Nudifloramide+LPC18:0
两个标记物的浓度都考虑(通过将Nudifloramide的浓度标准设为270pg/μl，将LPC18:0的浓度标准设为质谱仪au1,700,000)的确定结果如表4。下表中，“是”是指Nudifloramide低于270pg/μl和LPC 18:0的浓度超过1,700,000全都满足的情况，“否”是两个条件中不满足任何一个的情况。
[表4]
表4的确定结果表明，Nudifloramide低于270pg/μl和LPC 18:0超过1,700,000的胆道癌的灵敏度、特异性、PPV和NPV如下。
灵敏度：63.04％(58/92)
特异性：97.35％((28+337+99+30+58)/(34+340+100+30+63))
阳性预测值(PPV)：79.45％(58/(58+6+3+1+0+5))
阴性预测值(NPV):94.18％
((28+337+99+30+58)/(34+28+337+99+30+58))
如上面的结果将Nudifloramide和LPC18:0全都考虑时，胆道癌筛选的灵敏度有所减小，但特异性、PPV和NPV均增加，使其能够在实际临床中使用。
FAC
各种癌和正常对照组中的FAC的浓度分布如图5，以167pg/μl为基础的确定结果如表5。
[表5]
基于表5中的确定结果，基于FAC167pg/μl的胆道癌的灵敏度、特异性、PPV和NPV如下。
灵敏度：53.26％(49/92)
特异性：79.18％((11+321+34+25+58)/(34+340+100+30+63))
阳性预测值(PPV)：29.34％(49/(49+23+19+66+5+5))
阴性预测值(NPV):((11+321+34+25+58)/(43+11+321+34+25+58))
Nudifloramide+LPC18:0+FAC
Nudifloramide、LPC18:0和FAC全都考虑时的灵敏度、特异性、PPV和NPV如下表6。这里的标准浓度是Nudifloramide 270pg/μl，LPC18:0 1,700,000au和FAC 167pg/μl。
[表6]
如上面的结果把Nudifloramide、LPC18:0和FAC都考虑时，胆道癌筛选的灵敏度有所减小，但特异性、PPV和NPV均增加，使其能够在实际临床中使用。
上述的实施方案是为了说明本发明的示例，本发明并不限于此。本发明所属领域的技术人员应理解，在不背离随附权利要求限定的发明的精神和范围的情况下，可以在形式和细节方面做出各种改变。其它特征和方面将显而易见于下面的详细描述、附图和权利要求。

Claims (11)

1.一种用于提供针对胆道癌的诊断信息的方法，其特征在于，所述用于提供针对胆道癌的诊断信息的方法包括：
获取生物样品；
测量所述生物样品中用于预测胆道癌的标记物的浓度；以及使用测量的所述标记物的浓度提供针对胆道癌的诊断信息，其中所述标记物包括Nudifloramide。
2.根据权利要求1所述的用于提供针对胆道癌的诊断信息的方法，其特征在于，所述标记物还包括LPC18:0和纤维蛋白原α链中的至少一个。
3.根据权利要求1或2所述的用于提供针对胆道癌的诊断信息的方法，其特征在于，胆道癌诊断的Nudifloramide的标准浓度是220pg/μl至320pg/μl。
4.根据权利要求2所述的用于提供针对胆道癌的诊断信息的方法，其特征在于，胆道癌诊断用的所述纤维蛋白原α链的标准浓度是130pg/μl至200pg/μl。
5.根据权利要求2所述的用于提供针对胆道癌的诊断信息的方法，其特征在于，所述LPC18:0的浓度是利用质谱仪测定的，并且用于胆道癌诊断的所述LPC18:0的标准浓度是1,500,000au至2,000,000au。
6.根据权利要求2所述的用于提供针对胆道癌的诊断信息的方法，其特征在于，基于所述Nudifloramide和所述纤维蛋白原α链的特定浓度或更高浓度以及所述LPC18:0的特定浓度或更低浓度，所述信息的提供确定是否有胆道癌。
7.根据权利要求1所述的用于提供针对胆道癌的诊断信息的方法，其特征在于，所述生物样品包括血清。
8.根据权利要求1所述的用于提供针对胆道癌的诊断信息的方法，其特征在于，所述浓度通过质谱法测量。
9.一种用于诊断胆道癌的装置，其特征在于，所述用于诊断胆道癌的装置包括：输入单元，所述输入单元被构造为输入从生物样品中检测的质谱数据；以及诊断单元，所述诊断单元被构造为从所述质谱数据计算用于预测胆道癌的标记物的浓度，并基于计算的浓度确定针对胆道癌的诊断信息，其中所述标记物包括Nudifloramide。
10.根据权利要求9所述的用于诊断胆道癌的装置，其特征在于，所述标记物还包括LPC18:0和纤维蛋白原α链中的至少一个。
11.根据权利要求10所述的用于诊断胆道癌的装置，其特征在于，用于胆道癌诊断的所述Nudifloramide的标准浓度是220pg/μl至320pg/μl。
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