CN112305122A - 代谢物标志物及其在疾病中的应用 - Google Patents
代谢物标志物及其在疾病中的应用 Download PDFInfo
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- CN112305122A CN112305122A CN202011191207.3A CN202011191207A CN112305122A CN 112305122 A CN112305122 A CN 112305122A CN 202011191207 A CN202011191207 A CN 202011191207A CN 112305122 A CN112305122 A CN 112305122A
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Abstract
本发明公开了代谢物标志物及其在疾病中的应用,所述代谢物标志物为PC(O‑16:0/18:2(9Z,12Z)),所述疾病为动脉粥样硬化性脑梗死。在本发明的实施方案中,与动脉粥样硬化患者相比,代谢物标志物在动脉粥样硬化性脑梗死患者中水平降低。样本中该标志物的测量可表明受试者患有动脉粥样硬化性脑梗死或存在患动脉粥样硬化性脑梗死的风险。
Description
技术领域
本发明属于生物医药领域,涉及代谢物标志物及其在疾病中的应用。
背景技术
国民经济的快速发展让人民的生活质量得到了显著的改善,医疗技术水平的不断提高让人们的平均寿命获得了明显的延长,但也带了人口老龄化问题。脑血管病已经成为了威胁我国中老年人民身体健康和生命的主要疾病。目前,尚无有效手段能完全治愈该病,只能进行提前预防。脑血管病发病率的不断上升,给很多家庭带能来了困扰,给整个社会也造成了很大影响。所以,目前找出引起脑血管病的原因,并尽可能早的进行干预治疗,预防脑血管病发具有重要的意义。
脑梗死属于脑血管病的一种,该疾病是由很多原因共同影响而产生的,遗传因素是病因之一。高血压、糖尿病、高血脂以及动脉粥样硬化等都为脑血管病的危险因素,而动脉粥样硬化性脑梗死是脑梗死最常见的病理类型。目前,脑梗死的诊断主要依靠临床医师对症状、体征的判断及影像学检查,但是许多疾病如偏头痛、痫性发作等可能与脑梗死有类似的临床表现,影像学客观检查也有一定的局限性。在过去的数十年中,临床医生及科学家们致力于找到可以在脑梗死早期辅助诊断的生物标志物,利用生物标志物辅助临床医生在脑梗死早期做出诊断,将会使脑梗死患者及时得到有效救治,降低脑梗死的死亡率及病残率,在临床实践中意义重大。
代谢物组学是继基因组学和蛋白质组学之后新近发展起来的一门学科,是系统生物学的重要组成部分。其是对细胞、有机体分泌出来的体液进行动态跟踪分析,来辨识和解析被研究对象的生理、病理状态及其与环境因子、基因组成等的关系,已在临床诊断、药物开发、毒理学、生理学、病理学等研究领域得到了广泛的应用。通过代谢组学的方法检测疾病成为当前研究疾病的发生机理的一种重要手段。
发明内容
为了弥补现有技术的不足,本发明的目的在于提供与动脉粥样硬化性脑梗死相关的代谢物标志物,通过检测代谢物标志物的水平,可以判断患者是否患有动脉粥样硬化性脑梗死,从而为动脉粥样硬化性脑梗死的早期诊断提供一种新的手段。
术语“代谢物”是指代谢的中间产物和最终产物,(也有时称为具有小于1500道尔顿的分子量的小分子或分析物)。代谢物被分类为直接参与正常生长、发育和繁殖的初级代谢物。次级代谢物不直接参与后面的过程,但可以具有重要的生态学功能(例如,抗生素、色素)。代谢物的示例性生物学功能包括作为生物合成途径中的中间或终点产物或作为细胞信号分子。
为了实现上述目的,本发明提供了如下技术方案:
本发明的第一方面提供了一种与动脉粥样硬化性脑梗死相关的代谢物标志物,所述代谢物标志物包括PC(O-16:0/18:2(9Z,12Z))。
本发明的第二方面提供了检测样本中本发明第一方面所述的代谢物标志物水平的试剂在制备诊断动脉粥样硬化性脑梗死的产品中的应用。
进一步,所述试剂通过靶向或非靶向的核磁共振法、色谱法、光谱法、质谱法中的一种或几种检测样本中代谢物的水平。
进一步,所述色谱法包括气相色谱法、毛细管电泳法、液相色谱法、高度液相色谱法、超高效液相色谱法。
光谱法包括紫外可见光谱法、红外光谱法、近红外光谱法、核磁共振光谱法。
质谱法包括例如串联质谱法、基质辅助激光解吸电离(MALDI)飞行时间(TOF)质谱法、MALDI-TOF-TOF质谱法、MALDI四极杆-飞行时间(Q-TOF)质谱法、电喷射离子化(ESI)-TOF质谱法、ESI-Q-TOF、ESI-TOF-TOF、ESI-离子阱质谱法、ESI三重四极杆质谱法、ESI傅立叶变换质谱法(FTMS)、MALDI-FTMS、MALDI-离子阱-TOF和ESI-离子阱TOF。以其最基本的水平,质谱法涉及使分子电离和随后测量所得离子的质量。由于分子以公知的方式电离,可以精确地由离子的质量确定分子的分子量。
液相色谱质谱法结合了液相色谱法(LC)或高效液相色谱法(HPLC)的物理分离能力与质谱法(MS)的质量分析能力。HPLC提供超过LC的优点,其具有分析物的较短分析时间和较好分辨率。这因此增加了MS的选择性、精度和准确性。
串联质谱法涉及首先获得所关注的离子的质谱,随后破碎该离子并获得片段的质谱。串联质谱法因此提供分子量信息和破碎谱,其可一起结合分子量信息使用以识别肽或蛋白质或小分子(低于1500道尔顿)的确切序列。
进一步,所述试剂通过色谱-质谱法检测样本中代谢物的水平。
在本发明中,所述样本是生物学样本。生物来源的样本(即生物学样本)通常包含多种代谢物。待用于本发明方法的优选实验样本是来自体液,优选来自血液、血浆、血清、淋巴、汗、唾液、眼泪、精液、阴道液体、粪便、尿液或脑脊液的样本,或来自例如通过活体解剖来自细胞、组织或器官的样本。这也包括包含亚细胞区室或细胞器(如线粒体、高尔基体网络或过氧化物酶体)的样本。此外,生物学样本也包括气体样本,如生物体的挥发物。生物学样本来自如此处其他地方具体说明的受试者。用于获得上述不同类型生物学样本的技术为本领域所熟知。例如,通过血液采集获得血液样本,通过尿液采集获得尿液样本。
进一步,所述样本选自血液、血清、血浆。
进一步,在上述样本用于本发明检测前将其预处理。所述预处理可包括释放或分离化合物,或去除多余物质或废物所需要的处理。合适的技术包含离心、萃取、分馏、纯化和/或富集化合物。此外,进行其他预处理以提供适合于化合物分析的形式或浓度的化合物。例如,如果气相层析偶联质谱用于本发明的方法,将需要在所述气相层析前衍生化化合物。合适并必要的预处理依赖于进行本发明方法的工具并为本领域技术人员所熟知。如前描述的预处理的样本也包含在如本发明所用的术语“样本”中。
进一步,当受试者中的代谢物标志物的水平下调时,受试者患有动脉粥样硬化性脑梗死或者存在患动脉粥样硬化性脑梗死的风险。
在从已知未患有动脉粥样硬化性脑梗死的受试者或群体中获得参考结果的情况下,可以基于从试验样本中获得的试验结果与上述参考结果的差异,即基于关于至少一种代谢物的定性或定量组成中的差异来诊断所述疾病或易感性。差异可以是代谢物绝对或相对量的增加(有时称作代谢物上调)或代谢物所述量的减少或无可检测的量(有时称作代谢物下调)。优选地,相对或绝对量的差异是显著的,即在45到55百分位数、40到60百分位数、30到70百分位数、20到80百分位数、10到9百分位数、5到95百分位数的参考值区间外。相对量改变的优选值(即“倍数”变化)或改变类型(即导致更高或更低相对量和/或绝对量的“上”调或“下”调)。如果指定代谢物在受试者中是“上调的”,相对和/或绝对量将增加,如果其为“下调的”,代谢物的相对和/或绝对量将减少。此外,“倍数”变化表示增加或减少的程度,例如,2倍的增加指与参考相比,所述量是代谢物量的两倍。
因此,在优选实施方案中包括来自已知患有动脉粥样硬化性脑梗死的受试者或组的参考,或已知具有其易感性的受试者或组的参考。最优选地,试验样本和所述参考的相同或相似结果(即所述至少一种代谢物相似的相对或绝对量)在该情况下指示动脉粥样硬化性脑梗死或其易感性。在本发明另一个优选实施方案中,所述参考来自已知未患有动脉粥样硬化性脑梗死的受试者或已知不具有其易感性的受试者,或者所述参考是可计算参考。
本发明的第三方面提供了一种诊断动脉粥样硬化性脑梗死的试剂盒,所述试剂盒包括检测样本中本发明第一方面所述的代谢物标志物水平的试剂。
进一步,所述试剂通过靶向或非靶向的核磁共振法、色谱法、光谱法、质谱法中的一种或几种检测样本中代谢物的水平。
进一步,所述试剂盒还包含处理样本的试剂。
进一步,所述试剂盒还包括使用该试剂盒评估受试者是否患有动脉粥样硬化性脑梗死或存在患动脉粥样硬化性农安梗死的风险的说明书。
当在实验室环境中处理血液样本时,可能获得最可靠的结果。例如,可在医生办公室中从受试者获取血液样本,然后将其发送到医院或商业医学实验室进行进一步测试。然而,在许多情况下,可能希望在临床医生的办公室提供即时结果或允许受试者在家中进行测试。在一些情况下,对于便携式、预包装、一次性的、可由受试者在无协助或指导等的情况下即可使用等等的测试的需求比高度准确度更为重要。在许多情况下,尤其是在有医师随访的情况下,进行初步测试,甚至灵敏度和/或特异度降低的测试也可能就足够了。因此,以试剂盒形式提供的测定可涉及检测和测量相对少量的代谢物,以降低测定的复杂性和成本。
可使用本文所述的能够检测血液代谢物的任何形式的血液测定。通常,所述测定将定量血液代谢物至一定的程度,例如它们的浓度或量是高于还是低于预定阈值。此类试剂盒可采取测试条、浸杆、盒、药筒、基于芯片或基于珠粒的阵列、多孔板或一系列容器等的形式。提供一种或多种试剂以检测所选血液代谢物的存在和/或浓度和/或量。可将受试者的血液直接分配到测定中,或从存储的或先前获得的样品中间接分配到测定中。高于或低于预定阈值的代谢物的存在或不存在可以例如通过发色、发荧光、电化学发光或其他输出(例如如在酶免疫测定(EIA),诸如酶联免疫测定(ELISA)中)来显示。
在本发明中,试剂盒可包含固体基片诸如芯片、载玻片、阵列等,其具有能够检测和/或定量固定在基片上的预定位置处的一种或多种血液代谢物的试剂。作为说明性实例,可向芯片提供固定在离散的预定位置的试剂,以用于检测和定量血液样品中代谢物标志物,其任何数量或其任何组合的浓度。
本发明的第四方面提供了本发明第一方面所述的代谢物标志物在构建预测动脉粥样硬化性脑梗死的计算模型中的应用。
在本发明中,有许多方法可以评估选定的代谢物,以评估受试者是否患有或易患动脉粥样性硬化脑梗死。可将代谢物的测量值进行数学组合,并且可使组合后的值与潜在的状态问题发生关联。代谢物值可通过任何适当的数学方法组合。用于将代谢物或组合与疾病发生关联的数学方法可采用诸如但不限于判别分析(discriminant analysis,DA)(即线性、二次、正则化DA)、核方法(即SVM)、非参数方法(即k最近邻分类器)、PLS(偏最小二乘)、基于树的方法(即逻辑回归、CART、随机森林方法、增强/装袋方法)、广义线性模型(即对数回归)、基于主成分的方法(即SIMCA)、广义加性模型、基于模糊逻辑的方法、基于神经网络和遗传算法的方法之类的方法。对于SVM模型,可使用SVM分类器中每个特征的线性系数来选择最重要的特征。可选择具有最大绝对值的那些特征,并且可以根据需要仅使用所选特征和训练集来重新计算SVM模型。
当比较两个不同群体(例如,一个患病而另一个不患病)的测试结果时,很少观察到两组之间的完美分离。实际上,测试结果的分布会重叠,因此,当选择并应用区分两个群体的截断点或标准值时,在一些情况下该疾病会正确分类为阳性(真阳性分数),但一些疾病病例将归类为阴性(假阴性分数)。另一方面,一些没有疾病的病例将被正确地分类为阴性(真阴性分数),而一些没有疾病的病例将被分类为阳性(假阳性分数)。
可使用诸如ROC曲线分析之类的工具来评估这样的测试的性能,或测试将疾病组与对照组区分开的准确度。ROC曲线是使用灵敏度作为y轴,将1-特异性作为x轴,使用各种截断值生成的灵敏度和特异度谱图的图形表示。在ROC曲线中,针对不同的截断点绘制了真阳性率(灵敏度)与FP率(100-特异性)的函数关系。ROC曲线上的每个点代表对应于特定决策阈值的灵敏度/特异性对。具有完美判别力的测试(两个分布中没有重叠)的ROC曲线穿过左上角(灵敏度为100%,特异度为100%)。因此,从质量上讲,曲线图越靠近左上角,测试的总体准确度越高。ROC曲线(AUC)下面积反映了测试的准确度,并显示在曲线图的左下角。
本发明的优点和有益效果:
本发明首次发现了与动脉粥样硬化性脑梗死相关的代谢物标志物-PC(O-16:0/18:2(9Z,12Z)),通过检测上述代谢标志物的水平,可以判断受试者是否患有动脉粥样硬化性脑梗死以及患动脉粥样硬化性脑梗死的风险,以期实现动脉粥样硬化性脑梗死的早期的诊断,从而在动脉粥样硬化性脑梗死早期进行干预治疗,提高患者的生存质量。
附图说明
图1是OPLS-DA统计分析图,其中图A是反向色谱正离子统计分析图;图B是反向色谱负离子统计分析图;图C是亲水色谱正离子统计分析图。
图2是PC(O-16:0/18:2(9Z,12Z))在不同群组中的水平图。
图3是以PC(O-16:0/18:2(9Z,12Z))作为检测变量的诊断效能图。
具体实施方式
下面结合附图和实施例对本发明作进一步详细的说明。以下实施例仅用于说明本发明而不用于限制本发明的范围。实施例中未注明具体条件的实验方法,通常按照常规条件。
实施例动脉粥样硬化脑梗死相关代谢物的筛选及效能判断
1、样本收集
收集21例动脉粥样硬化脑梗死患者及21例动脉粥样硬化患者的血液样本。
动脉粥样硬化脑梗死组纳入标准:
1)受试者已签署知情同意书
2)符合《中国急性缺血性脑卒中诊疗指南(2014版)》急性脑梗死诊断标准。
3)病因分型为动脉粥样硬化型脑梗死。
4)年龄18-65周岁。
5)BMI 18.5-23.9kg/m2。
6)血常规:红细胞计数、MCHC、血红蛋白、白细胞计数、淋巴细胞计数、中性粒细胞计数、单核细胞计数在正常范围。
7)TG、TC、HDL-C、LDL-C、血糖、糖化血红蛋白在正常范围。
排除标准:
1)合并其他疾病:神经系统疾病(既往脑梗死、脑出血、多发性硬化等);各种慢性消化系统疾病,3个月内患有急性消化系统疾病;循环系统疾病(冠心病、心力衰竭、房颤);呼吸系统疾病(慢性阻塞性肺疾病、慢性支气管炎、哮喘);代谢性疾病(肥胖、高脂血症、糖尿病、代谢综合征、骨质疏松症);泌尿系统疾病(慢性肾脏病、肾衰竭、肾结石);血液系统疾病(贫血);其他(痛风、抑郁、精神疾病、慢性疲劳综合征、纤维肌痛症、食物过敏、肿瘤)。
2)既往有输血史、消化系统疾病手术史及外伤史。
3)心电图异常的患者。
4)3个月内服用以下药物:抗生素、泻药、氯硝西泮、性激素类药物、口服避孕药、美沙拉嗪、TNF-α抑制剂、免疫抑制剂、抗抑郁药、PPI、卢帕他定、阿片类、钙剂、维生素D、二甲双胍、叶酸、β-交感神经吸入剂、中药。
5)3个月内服用益生菌制剂。
6)本次发病前应用抗血小板及他汀类药物。
7)进行静脉溶栓和血管内介入治疗的患者。
8)妊娠期或哺乳期妇女。
9)本研究期间,患者已入选或计划入选另一项临床药物或装置/干预性研究。
动脉粥样硬化组纳入标准:
1)受试者已签署知情同意书。
2)颈部血管超声和/或颈部血管影像学表现为颅内外血管动脉粥样硬化。
3)年龄18-65周岁。
4)BMI 18.5-23.9kg/m2。
5)血常规:红细胞计数、MCHC、血红蛋白、白细胞计数、淋巴细胞计数、中性粒细胞计数、单核细胞计数在正常范围。
6)TG、TC、HDL-C、LDL-C、血糖、糖化血红蛋白在正常范围。
排除标准:
1)存在其他疾病:神经系统疾病(脑梗死、脑出血、多发性硬化等);各种慢性消化系统疾病,3个月内患有急性消化系统疾病;循环系统疾病(冠心病、心力衰竭、房颤);呼吸系统疾病(慢性阻塞性肺疾病、慢性支气管炎、哮喘);代谢性疾病(肥胖、高脂血症、糖尿病、代谢综合征、骨质疏松症);泌尿系统疾病(慢性肾脏病、肾衰竭、肾结石);血液系统疾病(贫血);其他(痛风、抑郁、精神疾病、慢性疲劳综合征、纤维肌痛症、食物过敏、肿瘤)。
2)既往有输血史、消化系统疾病手术史及外伤史。
3)心电图异常。
4)3个月内服用以下药物:抗生素、泻药、氯硝西泮、性激素类药物、口服避孕药、美沙拉嗪、TNF-α抑制剂、免疫抑制剂、抗抑郁药、PPI、卢帕他定、阿片类、钙剂、维生素D、二甲双胍、叶酸、β-交感神经吸入剂、中药、抗血小板药物及他汀类药物。
5)3个月内服用益生菌制剂。
6)妊娠或哺乳期妇女。
7)本研究期间,受试者已入选或计划入选另一项临床药物或装置/干预性研究。
2、非靶向代谢组学检测
2.1血清样本制备
2.1.1反相色谱分析血清样本处理方法
1)血浆/血清样本在4℃的冰融化为30-60min。
2)取40μl血清至标记好标签的1.5ml离心管中,加入300μl的甲醇和1ml甲基叔丁基醚。
3)充分振荡15s,进行蛋白沉淀。12000rpm,4℃,离心10min,取上层溶液100μl,置于200μl内衬管中,待测。
2.1.2亲水色谱分析血清样本处理方法:
1)血浆/血清样本在4℃的冰融化为30-60min。
2)取50μl血清至标记好标签的1.5ml离心管中,加入150μl的乙腈。
3)充分振荡15s,进行蛋白沉淀。12000rpm,4℃,离心10min,取上层溶液100μl,置于200μl内衬管中,待测。
2.2色谱条件
色谱分离采用Thermo Scientific的U3000快速液相色谱使用反相色谱和亲水色谱对血清样本进行分析。
2.2.1反相色谱分离条件
色谱柱:waters UPLC HSS T3(1.8μm 2.1mm*100mm);
流动相:A(乙腈/水4:6,0.1%甲酸,10mM乙酸铵)和B(乙腈/异丙醇9:1,0.1%甲酸,10mM乙酸铵);
洗脱程序:见表1;
流速:0.3ml/min;
进样量为1.0μL;
柱温:50℃。
表1 C18反相色谱测定洗脱程序
2.2.1亲水色谱分离条件
色谱柱:waters UPLC BEH Amide(1.7μm 2.1mm*100mm);
流动相:A(乙腈,0.1%甲酸,10mM乙酸铵)和B(水,0.1%甲酸,10mM乙酸铵);
洗脱程序:见表2;
流速:0.3ml/min;
进样量:1.0μL;
柱温:40℃。
表2 HILIC测定极性小分子洗脱程序
2.3质谱条件
质谱分析采用装备了热电喷雾离子源的四极杆轨道离子阱质谱仪。正负离子离子源电压分别为3.7kv和3.5kV。毛细管加热温度320℃。翘气压力30psi,辅助气压力10psi。容积加热蒸发温度300℃。翘气和辅助气均为氮气。碰撞气为氮气,压力为1.5mTorr。一级全扫描参数为:分辨率70000,自动增益控制目标为1×106,最大隔离时间50ms,质荷比扫描范围50-1500。液质系统由Xcalibur 2.2SP1.48软件控制,数据采集和靶向代谢物定量处理均由该软件操作。
3、靶向代谢组学检测
3.1血清样本处理方法
1)血浆样本于4℃放置30min解冻。
2)取50μl血浆样本至1.5ml离心管中,加入150μl的甲醇(含有吲哚乙酸-D2500ppb,吲哚丙酸-D2 50ppb组成),旋涡震荡30min。
3)12000rpm离心5min,取上清液100μl,置于200μl内衬管中,待测。
3.2色谱条件
色谱分离采用Waters ACQUITY UPLC I-CLASS超高压液相色谱系统,色谱分离条件如下:
色谱柱:Waters UPLC BEH C8(1.7μm 2.1mm*100mm);
流动相:A(水,0.5Mm NH4F)和B(甲醇);
洗脱梯度:见表3;
流速:0.3ml/min;
进样量:1.0μL;
柱温:45℃。
表3洗脱程序
3.3质谱条件
质谱分析仪器为Waters公司XEVO TQ-XS型串联四级杆质谱仪。正离子离子源电压为3kv,锥孔电压为20V。去溶剂温度550℃,源温度150℃。去溶剂气流速1000L/Hr,锥孔气流速7L/h。
3.4靶向代谢组数据处理
靶向代谢组数据峰面积计算采用masslynx定量软件,保留时间允许误差15s。浓度计算采用单点同位素内标法获取定量结果。
4、数据处理
4.1数据质控
为了评价样品采集过程中系统的稳定性和重复性,使用了质控样本。质控样本是所有样本均移取固定体积混合均匀后得到的。指控样本的前处理方法和其他样品一样。为了得到可信赖的且可重复性的代谢物,三个因素需要考虑:1)保留时间,2)信号强度,3)质量准确度。本次实验首先采用5个空白样本平衡色谱柱,再采用3个质控样本平衡柱条件。然后每间隔6-8个样本插入1个质控样本用于监测整个液质系统的稳定性和重复性。同时计算质控样本中提取的代谢特征的变异系数值,变异系数超过15%的代谢特征被删除。
4.2 PCA分析
所有采集好的数据,无论是何种分离模式或是正负离子模式,均采用ProgenesisQI软件处理,包括的步骤依次为导入原始数据、峰对齐、峰提取、归一化处理,最终形成保留时间、质荷比和峰强度的表格。反相色谱和亲水色谱提取峰的时间依次为1至16和1至12min。各种添加剂离子如加氢和加钠等均去卷积到每一个离子特征。代谢物鉴定采用人类代谢组数据库和脂质数据库进行一级分子量匹配。
4.3 OPLS-DA分析
为了获得在动脉粥样硬化性脑梗死组(BL)和在动脉粥样硬化组(AS)组呈现显著差异的代谢物信息,进一步采用监督性的多维统计方法即偏最小二乘方判别分析(OPLS-DA)对两组样本进行统计分析。
采用OPLS-DA模型的VIP(Variable Importance in the Projection)值(阈值>1),并结合t-test的p值(p<0.05)来寻找差异性表达代谢物。差异性代谢物的定性方法为:搜索在线数据库(HMDB)(比较质谱的质荷比m/z或者精确分子质量mass,误差限制0.01Da)。
4.4 ROC分析
根据代谢物的水平,绘制受试者工作特征曲线(ROC),计算二项精确置信空间,分析差异代谢物的诊断效能。
5、结果
质控结果显示,质控样本相对聚集在一起,系统重复性较好,所采集的数据可以进行进一步的研究。
反向色谱正离子、反向色谱负离子、亲水色谱正离子的结果分别如表4和图1所示。
表4 OPLS-DA分析模型参数
差异分析结果显示,与动脉粥样硬化组相比,PC(O-16:0/18:2(9Z,12Z))在动脉粥样硬化脑梗死组的水平显著降低(图2)。
以PC(O-16:0/18:2(9Z,12Z))的含量作为检测变量判断诊断效能,结果显示,曲线下面积为0.887,截断值为52754811.670,敏感性为0.952,特异性为0.810(图3),具有较高的敏感性和特异性。
上述实施例的说明只是用于理解本发明的方法及其核心思想。应当指出,对于本领域的普通技术人员来说,在不脱离本发明原理的前提下,还可以对本发明进行若干改进和修饰,这些改进和修饰也将落入本发明权利要求的保护范围内。
Claims (10)
1.一种与动脉粥样硬化性脑梗死相关的代谢物标志物,其特征在于,所述代谢物标志物包括PC(O-16:0/18:2(9Z,12Z))。
2.检测样本中权利要求1所述的代谢物标志物水平的试剂在制备诊断动脉粥样硬化性脑梗死的产品中的应用。
3.根据权利要求2所述的应用,其特征在于,所述试剂通过靶向或非靶向的核磁共振法、色谱法、光谱法、质谱法中的一种或几种检测样本中代谢物的水平。
4.根据权利要求3所述的应用,其特征在于,所述试剂通过色谱-质谱法检测样本中代谢物的水平。
5.根据权利要求2-4任一项所述的应用,其特征在于,所述样本选自血液、血清、血浆。
6.根据权利要求2-4任一项所述的应用,其特征在于,当受试者中的代谢物标志物的水平下调时,受试者患有动脉粥样硬化性脑梗死或者存在患动脉粥样硬化性脑梗死的风险。
7.一种诊断动脉粥样硬化性脑梗死的试剂盒,其特征在于,所述试剂盒包括检测样本中权利要求1所述的代谢物标志物水平的试剂。
8.根据权利要求7所述的试剂盒,其特征在于,所述试剂通过靶向或非靶向的核磁共振法、色谱法、光谱法、质谱法中的一种或几种检测样本中代谢物的水平。
9.根据权利要求7或8所述的试剂盒,其特征在于,所述试剂盒还包含处理样本的试剂。
10.权利要求1所述的代谢物标志物在构建预测动脉粥样硬化性脑梗死的计算模型中的应用。
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