CN112305120A - 代谢物在动脉粥样硬化性脑梗死中的应用 - Google Patents
代谢物在动脉粥样硬化性脑梗死中的应用 Download PDFInfo
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Abstract
本发明公开了代谢物在动脉粥样硬化性脑梗死中的应用。本发明通过代谢组学研究,首次发现了LysoPE(20:4(8Z,11Z,14Z,17Z)/0:0)在动脉粥样硬化脑梗死中呈现显著性差异,通过检测LysoPE(20:4(8Z,11Z,14Z,17Z)/0:0)的水平,可以判断受试者是否患有动脉粥样硬化脑梗死或者存在患有动脉粥样硬化性脑梗死的风险。
Description
技术领域
本发明属于生物技术领域,涉及代谢物在动脉粥样硬化性脑梗死中的应用。
背景技术
脑梗死又称缺血性卒中,中医称之为卒中或中风。在目前临床实践中,卒中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、痫性发作等可能与卒中有类似的临床表现,影像学客观检查也有一定的局限性。卒中包括出血性及缺血性两种类型,其中,动脉粥样硬化性脑卒中是缺血性脑卒中最常见的病理类型。疾病早期,颅脑电子计算机断层扫描(computed tomography,CT)是鉴别脑梗死与脑出血的唯一依据,CT早期诊断脑出血的敏感性为89%,但是在脑梗死患者的CT检查中,仅有不到1/3的患者在发病3h内有相应的特征性表现,磁共振成像(magnetic resonanceimaging,MRI)诊断脑梗死敏感性较高,但受到检查时间及设备等限制,不适用于急诊脑梗死患者。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,虽然目前尚无一种或几种有明确指导意义的生物标记物可以用于脑梗死的辅助诊断,但是随着气相色谱-质谱法、液相色谱-质谱法、基因芯片、蛋白质芯片等科学技术的不断进步,越来越多与脑梗死相关的诊断性生物标记物被发现。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。
代谢组学是研究生物体系受刺激或扰动后其代谢产物一内源性代谢物质种类、数量及其变化规律的科学。生命机体在病证状态和各类药物作用下,亦同样会引起全身水平的内源性代谢物及代谢网络变化。运用代谢组学技术考察和分析这些代谢物的变化,对于探究病证本质,阐明药物作用机理,实现疾病的早期诊断和治疗具有重要的意义。
发明内容
为了弥补现有诊治技术的不足,本发明的目的在于提供一种与动脉粥样硬化脑梗死相关的代谢物,通过检测该代谢物的水平,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,甚至,采用药物后,疾病是否好转,从而为动脉粥样硬化性脑梗死的诊断和治疗提供新的手段。
本发明提供了LysoPE(20:4(8Z,11Z,14Z,17Z)/0:0)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,所述产品包括通过色谱、光谱或质谱仪/光谱法进行检测样品中LysoPE(20:4(8Z,11Z,14Z,17Z)/0:0)含量的试剂。
进一步,所述色谱包括GC,CE,LC,HPLC和UHPLC;光谱包括UV/Vis,IR,NIR和NMR;质谱仪/光谱法包括ESI、四极质谱仪、离子阱质谱仪、TOF(飞行时间)质谱仪、Orbitrap质谱仪、扇形磁场质谱仪、扇形静电场质谱仪、离子回旋共振(ICR)以及它们的组合,包括单级四极杆(Q)和三级四极杆(QqQ),QqTOF,TOF-TOF,Q-Orbitrap,APCI-QqQ,APCI-QqTOF,MALDI-QqQ,MALDI-QqTOF和MALDI-TOF-TOF。
进一步,所述样品选自血液、血清或血浆。
本发明提供了一种检测动脉粥样硬化性脑梗死的试剂盒,所述试剂盒包括检测样品中LysoPE(20:4(8Z,11Z,14Z,17Z)/0:0)含量的试剂。
进一步,所述试剂包括通过色谱、光谱或质谱仪/光谱法检测样品中LysoPE(20:4(8Z,11Z,14Z,17Z)/0:0)含量的试剂。
进一步,所述试剂盒还包含处理样品的试剂。
进一步,所述样品选自血液、血清或血浆。
进一步,所述试剂盒还包括评估受试者是否患有或易患动脉粥样硬化性脑梗死的说明书。
本发明提供了LysoPE(20:4(8Z,11Z,14Z,17Z)/0:0)在构建预测动脉粥样硬化性脑梗死的计算模型中的应用。
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-LysoPE(20:4(8Z,11Z,14Z,17Z)/0:0),通过检测样品中LysoPE(20:4(8Z,11Z,14Z,17Z)/0:0)的水平,可以判断受试者是否患有动脉粥样硬化性脑梗死或者患动脉粥样硬化性脑梗死的风险。
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LysoPE(20:4(8Z,11Z,14Z,17Z)/0:0)在不同群组中的含量图。
图3是以LysoPE(20:4(8Z,11Z,14Z,17Z)/0:0)作为检测变量的诊断效能图。
具体实施方式
代谢组学是基因组学、蛋白质组学和转录组学下游的新兴研究领域。人体中有40,000多种代谢物,其浓度可提供个体当前健康状况的快照。代谢组是由代谢产生的低分子量化合物(诸如代谢底物和产物、脂质、小肽、维生素和其它蛋白质辅助因子)的定量集合。代谢组处于转录组和蛋白质组的下游,因此从正常状态发生的任何变化都会被放大,并且在数值上更易于处理。代谢组学可以是检查和描述细胞生长、维持和功能的精确、一致和定量的工具。
为了评估代谢物与动脉粥样硬化性脑梗死之间的相关性,通过收集动脉粥样硬化患者与动脉粥样硬化性脑梗死的样品,综合分析样品的代谢组学,筛选在两个群组中含量呈现显著性差异的代谢物,并进一步分析差异代谢物的诊断效能,从而发现适于动脉粥样硬化性脑梗死诊断和治疗的代谢标志物。本发明通过广泛而深入的研究,首次发现了与动脉粥样硬化性脑梗死相关的代谢标志物LysoPE(20:4(8Z,11Z,14Z,17Z)/0:0),与动脉粥样硬化患者相比,LysoPE(20:4(8Z,11Z,14Z,17Z)/0:0)在动脉粥样硬化性脑梗死患者中含量显著增加。
本文使用的术语“代谢标志物”、“代谢生物标志物”或短的“生物标志物”被定义为适合作为动脉粥样硬化性脑梗死存在和状态的指标化合物,这种化合物是哺乳动物体内的代谢过程中出现的代谢物或代谢化合物。术语“生物标志物”和“代谢生物标志物”通常在本发明的上下文中同义使用,并且通常是指一种代谢物的量或两种或更多种代谢物的含量或比率。因此,术语“代谢生物标志物”或“生物标志物”也包括两种或更多种代谢物之间的含量或比率。
本文所用术语“差异代谢物”或“显著性差异”表示与第二种样品中相同的一种或多种本发明生物标志物的表达水平比较,经测定代谢物的含量或浓度,在一个样品中本发明的一种或多种生物标志物的含量或浓度的差异。本文所用“差异代谢物”可以用给定生物标志物的水平相对于对照中给定生物标志物的平均水平的比率进行测定,其中比率不等于1.0。还可以用p值测定差异。当使用p值时,当p值小于0.1时生物标志物被鉴定为在第一和第二群体之间呈现差异。更优选p值小于0.05。甚至更优选p值小于0.01。还更优选p值小于0.005。最优选p值小于0.001。当基于比率确定差异时,如果第一种和第二种样品中水平的比率大于或小于1.0,则代谢物是呈现差异的。举例来说,大于1.2、1.5、1.7、2、3、4、10、20的比率,或小于1的比率,例如0.8、0.6、0.4、0.2、0.1、0.05。
“水平增加”或“上调”表示代谢物相对于对照而言,代谢物水平(以代谢物的含量或浓度测定)显示增加至少10%或更多,例如20%、30%、40%或50%、60%、70%、80%、90%或更多;或1.1倍、1.2倍、1.4倍、1.6倍、1.8倍或更多。
“水平降低”或“下调”表示代谢物相对于对照而言,代谢物的水平(以代谢物的含量或浓度测定)显示降低至少10%或更多,例如20%、30%、40%或50%、60%、70%、80%、90%;或少于1.0倍、0.8倍、0.6倍、0.4倍、0.2倍、0.1倍或更少。例如,上调代谢物包括与从动脉粥样硬化患者中检测的代谢物的水平相比,从以患有动脉粥样硬化性脑梗死的个体中检测的水平增加的代谢物。例如,下调代谢物包括与从动脉粥样硬化患者中检测的代谢物的水平相比,从以患有动脉粥样硬化性脑梗死的个体中检测的水平减少的代谢物。
本发明技术人员应当理解,可以利用本领域内已知的任何方法来测定代谢物的水平,例如色谱法、光谱法和质谱法,而质谱法是特别优选的。色谱可以包括GC,LC,HPLC和UHPLC;光谱可包括UV/Vis,IR和NMR;质谱分析器/光谱可以包括ESI-QqQ,ESI-QqTOF,MALDI-QqQ,MALDI-QqTOF和MALDI-TOF-TOF。更优选地,质量分析器/光谱分析包括四极杆质量分析器,离子阱质谱分析器,TOF(飞行时间)质量分析器、轨道阱质量分析器、磁式扇形质量分析器、静电场扇形质量分析器(Electrostatic Sector Mass Analyzer)、离子回旋共振(ICR)以及质量分析器的组合(包括单四极杆(Q)和三重四极杆(QqQ),QqTOF,TOF-TOF,Q轨道阱)。优选是使用FLA-和HPLC-串联质谱法。
其中,GC=气相色谱,CE=毛细管电泳,LC=液相色谱,HPLC=高度液相色谱,UHPLC=超高效液相色谱,UV-Vis=紫外可见,IR=红外,NIR=近红外,NMR=核磁共振,ESI=电子喷雾电离,MALDI=基质辅助激光解吸/电离,TOF=飞行时间,APCI=大气压化学电离,QqQ=三重四极杆配置(也称为Qlq2Q3(Q1和Q3四极杆是质量过滤器,q2是无质量分辨四极杆(no mass-resolving quadrupole)))。
尽管使用一种代谢物对于诊断动脉粥样硬化性脑梗死疾病就足够了,但使用一种或多种,两种或多种,三种或多种,或四种或多种这样的代谢物包括在本发明内,并且在许多情况中,这是优选的。可以分析代谢物,并且以任意组合用于诊断。
术语“曲线下面积”或“AUC”指受试者工作特征(ROC)曲线的曲线下面积,二者是本领域中熟知的。AUC测量对于跨全部数据范围比较分类器的准确度是有用的。具有较高AUC的分类器具有在两个目标组(例如,动脉粥样硬化性脑梗死样品和动脉粥样硬化样品中)之间进行正确分类未知的更高的能力。ROC曲线对于在两个群体(例如,对动脉粥样硬化性脑梗死样品和动脉粥样硬化的个体)之间进行区别时描绘特定特征(例如,本文所描述的任何生物标志物和/或另外的生物医学信息的任何条目)的性能是有用的。通常,以单个特征的值为基础以升序顺序跨越整个群体(例如,病例和对照)选出特征数据。然后,对于该特征的每个值,计算数据的真阳性和假阳性率。真阳性率通过计数高于该特征的值的病例的数目并除以病例总数而确定。假阳性率通过计数高于该特征的值的对照的数目并除以对照总数而确定。虽然该定义指与对照相比较在病例中特征是增高的情况,该定义还应用于与对照相比较在病例中特征较低的情况(在该情况中,将计数低于该特征的值的样品)。ROC曲线可关于单独特征来生成,且可关于其他单独输出来生成,例如,两个或更多个特征的组合可用数学方法结合(例如,相加、相减、相乘等)以提供单独的总和值,且该单独的总和值可绘制于ROC曲线中。另外,其中的组合源自单独的输出值的多个特征的任意组合可绘制于ROC曲线中。特征的这些组合可包括试验。ROC曲线是试验的真阳性率(敏感性)针对试验的假阳性率(1-特异性)的图。
属于“样品”取自哺乳动物,优选为小鼠,大鼠,豚鼠,狗,小型猪或人,最优选为人。任何含有目标代谢物的生物样品在本文中是有用的。实例包括,但不限于,血液/血清/血浆、脑脊髓液(CSF)、尿液、粪便、呼气、唾液或任何组织的活检样品。优选的,所述样品选自血液。优选的,所述样品选自血清。
对于样品中一种或多种代谢物的量的评估,可能需要根据样品的类型和选择用于评估的方法操作样品。例如,在血液的情况下,可以将血液抽入合适的容器中,随后一次或多次平缓颠倒容器,并且让样品在室温下静止数分钟以获得完全凝固。对于血清采集,可以例如以2750g和15℃进行血液的离心10分钟。随后可以分离血清,并且如果需要,填充到容器例如合成吸管中,用于存储,如在液氮中,直到进行代谢分析。
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 2.2 SP1.48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);
洗脱程序: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。质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
生物信息学分析结果显示,与动脉粥样硬化组相比,LysoPE(20:4(8Z,11Z,14Z,17Z)/0:0)在动脉粥样硬化脑梗死组的水平显著增加(图2)。
以LysoPE(20:4(8Z,11Z,14Z,17Z)/0:0)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0.807,截断值为587403.738,敏感性为0.762,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.LysoPE(20:4(8Z,11Z,14Z,17Z)/0:0)在制备诊断动脉粥样硬化性脑梗死的产品中的应用。
2.根据权利要求1所述的应用,其特征在于,所述产品包括通过色谱、光谱或质谱仪/光谱法进行检测样品中LysoPE(20:4(8Z,11Z,14Z,17Z)/0:0)含量的试剂。
3.根据权利要求2所述的应用,其特征在于,所述色谱包括GC,CE,LC,HPLC和UHPLC;光谱包括UV/Vis,IR,NIR和NMR;质谱仪/光谱法包括ESI、四极质谱仪、离子阱质谱仪、TOF(飞行时间)质谱仪、Orbitrap质谱仪、扇形磁场质谱仪、扇形静电场质谱仪、离子回旋共振(ICR)以及它们的组合,包括单级四极杆(Q)和三级四极杆(QqQ),QqTOF,TOF-TOF,Q-Orbitrap,APCI-QqQ,APCI-QqTOF,MALDI-QqQ,MALDI-QqTOF和MALDI-TOF-TOF。
4.根据权利要求2或3所述的应用,其特征在于,所述样品选自血液、血清或血浆。
5.一种检测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品中LysoPE(20:4(8Z,11Z,14Z,17Z)/0:0)含量的试剂。
6.根据权利要求5所述的试剂盒,其特征在于,所述试剂包括通过色谱、光谱或质谱仪/光谱法检测样品中LysoPE(20:4(8Z,11Z,14Z,17Z)/0:0)含量的试剂。
7.根据权利要求6所述的试剂盒,其特征在于,所述试剂盒还包含处理样品的试剂。
8.根据权利要求5-7任一项所述的试剂盒,其特征在于,所述样品选自血液、血清或血浆。
9.根据权利要求5所述的试剂盒,其特征在于,所述试剂盒还包括评估受试者是否患有或易患动脉粥样硬化性脑梗死的说明书。
10.LysoPE(20:4(8Z,11Z,14Z,17Z)/0:0)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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