CN107677825A - 一种用于***诊断的肿瘤诊断组合物和用于制备诊断试剂盒的用途 - Google Patents

一种用于***诊断的肿瘤诊断组合物和用于制备诊断试剂盒的用途 Download PDF

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CN107677825A
CN107677825A CN201710926283.6A CN201710926283A CN107677825A CN 107677825 A CN107677825 A CN 107677825A CN 201710926283 A CN201710926283 A CN 201710926283A CN 107677825 A CN107677825 A CN 107677825A
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杨豪伟
刘义俊
王斌
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Han's joint (Tianjin) Stem Cell Research Institute Co., Ltd.
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Abstract

本发明公开了一种用于***诊断的肿瘤诊断组合物和用于制备诊断试剂盒的用途。单独使用血清溶血磷脂酰胆碱(18:2)、磷脂酰肌醇(20:4/0:0)或N‑苯丙酰‑L‑谷氨酰胺诊断区分***患者与健康志愿者时，ROC曲线下面积(AUC)均低于0.7；三者联合用于诊断区分***患者与健康志愿者时AUC可以提高到0.945。本领域技术人员知道，ROC曲线下面积在1.0和0.5之间，在AUC>0.5的情况下，AUC越接近于1，说明诊断效果越好。AUC在0.5‑0.7时有较低准确性，AUC在0.7‑0.9时有一定准确性，AUC在0.9以上时有较高准确性。因此，血清溶血磷脂酰胆碱(18:2)、磷脂酰肌醇(20:4/0:0)和N‑苯丙酰‑L‑谷氨酰胺可以联合用于制备诊断***的诊断试剂盒。

Description

一种用于***诊断的肿瘤诊断组合物和用于制备诊断试剂 盒的用途
技术领域
本发明属于疾病诊断领域，涉及诊断标志物，具体涉及一种用于***诊断的肿瘤诊断组合物和用于制备诊断试剂盒的用途。
背景技术
***是女性最常见的生殖道恶性肿瘤之一，高居女性生殖***癌症的首位，近年来，随着人类***状病毒等性传播疾病的增加，***的发病率逐年升高，且发病年龄趋向年轻化。***早期治疗预后良好，而晚期发现则预后极差。因此，***的早发现、早诊断、早治疗对提高***患者的生存率意义重大。
近年来，用组学技术来研究发病及病程机制的报道逐渐增多，包括基因组学、MicroRNA组学、蛋白组学、代谢组学等。其中，代谢组学能够跟踪检测代谢物之间的相互转化与含量水平变化，这些信息与病理生理过程中的生物化学以及生理功能改变联系起来可以作为潜在靶标和作用位点，进而确定相关的生物标志物。与基因组学和蛋白组学等其他组学技术相比较，代谢组学可以展现其优势：①代谢组反应基因和蛋白层面综合作用下的最终结果，而基因和蛋白的微弱变化在代谢组中具有“延增效应”；②机体中小分子化合物的组成相对简单，更容易阐释其生物学功能；③代谢组的数据分析可通过KEGG或HMDB等数据库进行辅助。
代谢标志物用于诊断***可以实现***诊断的无创化、高效化和人性化。
发明内容
本发明的目的在于提供一种用于***诊断的肿瘤诊断组合物和用于制备诊断试剂盒的用途，以通过血清就可以快速诊断***。
本发明通过下面的技术方案得以实现：
溶血磷脂酰胆碱(18:2)、磷脂酰肌醇(20:4/0:0)和N-苯丙酰-L-谷氨酰胺联合用于制备诊断***的诊断组合物的用途。
优选地，所述溶血磷脂酰胆碱(18:2)、磷脂酰肌醇(20:4/0:0)和N-苯丙酰-L-谷氨酰胺为血清溶血磷脂酰胆碱(18:2)、血清磷脂酰肌醇(20:4/0:0)和血清N-苯丙酰-L-谷氨酰胺。
溶血磷脂酰胆碱(18:2)、磷脂酰肌醇(20:4/0:0)和N-苯丙酰-L-谷氨酰胺联合用于制备诊断***的诊断试剂盒的用途。
优选地，所述溶血磷脂酰胆碱(18:2)、磷脂酰肌醇(20:4/0:0)和N-苯丙酰-L-谷氨酰胺为血清溶血磷脂酰胆碱(18:2)、血清磷脂酰肌醇(20:4/0:0)和血清N-苯丙酰-L-谷氨酰胺。
本发明的优点：
单独使用血清溶血磷脂酰胆碱(18:2)、磷脂酰肌醇(20:4/0:0)或N-苯丙酰-L-谷氨酰胺诊断区分***患者与健康志愿者时，ROC曲线下面积(AUC)均低于0.7；三者联合用于诊断区分***患者与健康志愿者时AUC可以提高到0.945。本领域技术人员知道，ROC曲线下面积在1.0和0.5之间，在AUC>0.5的情况下，AUC越接近于1，说明诊断效果越好。AUC在0.5-0.7时有较低准确性，AUC在0.7-0.9时有一定准确性，AUC在0.9以上时有较高准确性。因此，血清溶血磷脂酰胆碱(18:2)、磷脂酰肌醇(20:4/0:0)和N-苯丙酰-L-谷氨酰胺可以联合用于制备诊断***的诊断试剂盒。
附图说明
图1为血清溶血磷脂酰胆碱(18:2)单独用于区分***患者与健康志愿者的ROC曲线；
图2为血清磷脂酰肌醇(20:4/0:0)单独用于区分***患者与健康志愿者的ROC曲线；
图3为血清N-苯丙酰-L-谷氨酰胺单独用于区分***患者与健康志愿者的ROC曲线；
图4为血清溶血磷脂酰胆碱(18:2)、磷脂酰肌醇(20:4/0:0)和N-苯丙酰-L-谷氨酰胺联合用于区分***患者与健康志愿者的ROC曲线。
具体实施方式
下面结合具体实施例进一步介绍本发明的技术方案。
一、实验材料
***患者血清标本取自东南大学附属中大医院***手术后患者。所有患者术前均未接受放射治疗、化学治疗及抗肿瘤药物等治疗，均有完整的临床及病理资料。***患者95例(***组)，年龄31-62岁，平均年龄53.3岁。
健康志愿者血清标本40例(正常对照组)，年龄32-65岁，平均年龄54.5岁。
***组和正常对照组间年龄构成差异无统计学意义。
二、实验方法
1、血清样本前处理
取200μL血清于1.5mL离心管中，加入50μL 1mg/mL的2-异丙基苹果酸溶液内标，涡旋20秒混匀，加入400μL甲醇、氯仿和水的混合溶液(比例为2.5∶1∶1)，然后在70℃的金属浴上振摇30min(1200rpm)，16000g×5min离心(4℃)，取500μL上清于1.5mL离心管中，加入500μL蒸馏水，涡旋混匀，然后16000g×5min离心(4℃)，取500μL上清于1.5mL离心管中，在室温下用氮吹仪吹干，所得残渣用80μL的甲氧胺吡啶溶液溶解，在50℃条件下肟化8h，加入60μLN-甲基-N-三甲基硅基三氟乙酰胺，在70℃条件下衍生化2h，即得。
2、GC-MS分析
取上述衍生化的样品2μL进行GC-MS分析。
液相色谱条件如下：进样口温度：270℃；进样量：2.0μL；不分流进样，载气：高纯氦(99.999％)；流速：1.0mL/min；接口温度：260℃；色谱柱：DB-5MS毛细管柱，色谱柱程序升温进行检测，80℃恒温2min，80℃-300℃(5℃/min)恒温6min。
质谱条件如下：离子源温度：230℃；四级杆温度：150℃，溶剂延迟：5min；电子碰撞电离电压：70eV，质谱全扫描范围(m/z)：30-600，采用全扫描方式。
分别用溶血磷脂酰胆碱(18:2)、磷脂酰肌醇(20:4/0:0)或N-苯丙酰-L-谷氨酰胺的色谱峰面积除以内参2-异丙基苹果酸的峰面积，所得比值分别作为各样本中溶血磷脂酰胆碱(18:2)、磷脂酰肌醇(20:4/0:0)或N-苯丙酰-L-谷氨酰胺的相对含量。
3、统计学处理
采用SPSS 20.0软件进行分析，以均值±偏差表示，组间比较采用t检验。评价各项指标应用价值，绘制受试者工作特征(ROC)曲线，并计算曲线下面积(AUC)；以P＜0.05为差异有统计学意义。采用ROC曲线评价各单项指标及联合指标对***的诊断价值。
三、实验结果
1、两组间肿瘤标志物水平的比较
***组血清中溶血磷脂酰胆碱(18:2)的相对含量明显低于正常对照组，***组血清中磷脂酰肌醇(20:4/0:0)、N-苯丙酰-L-谷氨酰胺的相对含量均明显高于正常对照组，差异均有统计学意义(t值分别为1.733、1.479和2.314，P＜0.01)。比较结果如表1所示。
表1两组间肿瘤标志物水平的比较(均值±偏差)
2、肿瘤标志物检测***ROC曲线分析
单独使用血清溶血磷脂酰胆碱(18:2)、磷脂酰肌醇(20:4/0:0)或N-苯丙酰-L-谷氨酰胺诊断区分***患者与健康志愿者时，ROC曲线下面积(AUC)均低于0.7；三者联合用于诊断区分***患者与健康志愿者时AUC可以提高到0.945。由此可见，血清溶血磷脂酰胆碱(18:2)、磷脂酰肌醇(20:4/0:0)和N-苯丙酰-L-谷氨酰胺三者联合用于诊断***具有较高的临床价值。三个指标的ROC曲线分析结合二元逻辑回归操作。
ROC曲线分析结果如表2和图1-4所示。
表2三个血清代谢物单独及联合诊断价值比较
本领域技术人员知道，ROC曲线下面积在1.0和0.5之间，在AUC>0.5的情况下，AUC越接近于1，说明诊断效果越好。AUC在0.5-0.7时有较低准确性，AUC在0.7-0.9时有一定准确性，AUC在0.9以上时有较高准确性。因此，血清溶血磷脂酰胆碱(18:2)、磷脂酰肌醇(20:4/0:0)和N-苯丙酰-L-谷氨酰胺可以联合用于制备诊断***的诊断试剂盒。

Claims (4)

1.溶血磷脂酰胆碱(18:2)、磷脂酰肌醇(20:4/0:0)和N-苯丙酰-L-谷氨酰胺联合用于制备诊断***的诊断组合物的用途。
2.根据权利要求1所述的用途，其特征在于：所述溶血磷脂酰胆碱(18:2)、磷脂酰肌醇(20:4/0:0)和N-苯丙酰-L-谷氨酰胺为血清溶血磷脂酰胆碱(18:2)、血清磷脂酰肌醇(20:4/0:0)和血清N-苯丙酰-L-谷氨酰胺。
3.溶血磷脂酰胆碱(18:2)、磷脂酰肌醇(20:4/0:0)和N-苯丙酰-L-谷氨酰胺联合用于制备诊断***的诊断试剂盒的用途。
4.根据权利要求2所述的用途，其特征在于：所述溶血磷脂酰胆碱(18:2)、磷脂酰肌醇(20:4/0:0)和N-苯丙酰-L-谷氨酰胺为血清溶血磷脂酰胆碱(18:2)、血清磷脂酰肌醇(20:4/0:0)和血清N-苯丙酰-L-谷氨酰胺。
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