CN103952465A - 一种基于血浆microRNA分析的卵巢癌肿瘤诊断方法 - Google Patents

一种基于血浆microRNA分析的卵巢癌肿瘤诊断方法 Download PDF

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CN103952465A
CN103952465A CN201310698673.4A CN201310698673A CN103952465A CN 103952465 A CN103952465 A CN 103952465A CN 201310698673 A CN201310698673 A CN 201310698673A CN 103952465 A CN103952465 A CN 103952465A
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ovarian cancer
plasma
blood plasma
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micrornas
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易银沙
张万明
曹蓬荣
邹义洲
李娜
刘彩云
袁炳秋
吴小宁
冯志明
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CHANGSHA YINGRUN BIOTECHNOLOGY Co Ltd
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Abstract

一种基于血浆microRNA分析的卵巢癌肿瘤诊断方法，包括以下步骤：建立血浆microRNAs抽提、qRT-PCR定量检测的技术平台；分析卵巢癌患者和正常志愿者血浆microRNAs表达芯片，寻找差异表达的microRNAs；筛选血浆样本中肿瘤特异性高表达的miRNAs作为候选血浆miRNAs标志物；在卵巢癌患者的血浆中，通过检测血浆中特异高表达的miRNAs含量的变化，结合医学统计学，初步评价与确定了候选血浆miRNAs诊断卵巢癌的能力。本发明提供血浆提取miRNA诊断卵巢癌的新型方法、利用基因重组技术制备特异性miRNA的方法，本方法更方便，更灵敏，成本更低。

Description

一种基于血浆microRNA分析的卵巢癌肿瘤诊断方法
技术领域
本发明涉及血浆组织样本的提取与检测方法，具体为一种基于血浆microRNA分析的卵巢癌肿瘤诊断方法。 
背景技术
卵巢癌是卵巢肿瘤的一种恶性肿瘤，是指生长在卵巢上的恶性肿瘤，其中90％～95％为卵巢原发性的癌，另外5％～10％为其它部位原发的癌转移到卵巢。卵巢癌也是女性生殖器官常见的肿瘤之一发病率仅次于子宫颈癌和子宫体癌而列居第三位。但因卵巢癌致死者，却占各类妇科肿瘤的首位对妇女生命造成严重威胁。 
由于卵巢癌早期缺少症状，最初常无症状，部分病人无意中摸到下腹部包块或妇科检查时偶然发现。可常感下腹部不适，一般无明显腹痛。当出现并发症如蒂扭转、破裂、感染时可出现下腹部疼痛。部分病人可出现月经失调或闭经。即使有症状也不特异，筛查的作用又有限，因此早期诊断比较困难，就诊时60％～70％已为晚期，而晚期病例又疗效不佳。因此，虽然卵巢癌的发病率低于宫颈癌和子宫内膜癌居妇科恶性肿瘤的第三位，但死亡率却超过宫颈癌及子宫内膜癌之和，高居妇科癌症首位，是严重威胁妇女健康的最大疾患。因此对于卵巢癌的早期诊断是一个亟待解决的问题。 
发明内容
本发明所解决的技术问题在于提供一种基于血浆microRNA分析的卵巢癌肿瘤诊断方法，以解决上述背景技术中提出的问题。 
本发明所解决的技术问题采用以下技术方案来实现： 
一种基于血浆microRNA分析的卵巢癌肿瘤诊断方法，包括以下步骤： 
(1)建立血浆microRNAs抽提、qRT-PCR定量检测的技术平台； 
(2)利用miRNAsmicroarray分析卵巢癌患者和正常志愿者血浆microRNAs表达芯片，寻找差异表达的microRNAs(参照序列1)； 
(3)利用Northern Blot或qRT-PCR技术(添加的miRNA-QPCR引物序列为序列2)，筛选血浆样本中肿瘤特异性高表达的miRNAs作为候选血浆miRNAs标志物； 
(4)在卵巢癌患者的血浆中，通过检测血浆中特异高表达的miRNAs含量的变化，结合医学统计学，初步评价与确定了候选血浆miRNAs诊断卵巢癌的能力。 
其中，序列1为用于卵巢癌诊断的miRNA序列，序列1中miR-200a，miR-100，miR-141，miR-200b，and miR-200c等为miRNA特征性编码核酸序列以及其对应的RNA序列，以及其剪切之后的成熟序列。 
序列2中 
用于特异性扩增：miR-200a，miR-100，miR-141，miR-200b，and miR-200c的Q-PCR的引物。 
有益效果 
本发明提供血浆提取miRNA诊断卵巢癌的新型方法、利用基因重组技术制备特异性miRNA的方法，本方法更方便，更灵敏，成本更低。 
附图说明
图1为正常对照组血清样品的miRNAl41c与对照miRNA(a)变化标准曲线； 
图2为卵巢癌患者中血清样品的miRNA141c与对照miRNA(a)变化标准曲线。 
具体实施方式
下面将结合本发明实施例，对本发明实施例中的技术方案进行清楚、完整地描述，显然，所描述的实施例仅是本发明一部分实施例，而不是全部的实施例。基于本发明中的实施例，本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例，都属于本发明保护的范围。 
实施例1 
一种基于血浆microRNA分析的卵巢癌肿瘤诊断方法，包括以下步骤： 
(1)建立血浆microRNAs抽提、qRT-PCR定量检测的技术平台； 
(2)利用miRNAsmicroarray分析卵巢癌患者和正常志愿者血浆microRNAs表达芯片，寻找差异表达的microRNAs； 
(3)利用Northern Blot或qRT-PCR技术，筛选血浆样本中肿瘤特异性高表达的miRNAs作为候选血浆miRNAs标志物； 
(4)在卵巢癌患者的血浆中，通过检测血浆中特异高表达的miRNAs含量的变化，(如图1-2所示)结合医学统计学，初步评价与确定了候选血浆miRNAs诊断卵巢癌的能力。 
通过大规模的样品分析确定miRNA在卵巢肿瘤诊断上的运用，可以进行相对应的试剂盒的研发与市场开发。 
对于本领域技术人员而言，显然本发明不限于上述示范性实施例的细节，而且在不背离本发明的精神或基本特征的情况下，能够以其他的具体形式实现本发明。因此，无论从哪一点来看，均应将实施例看作是示范性的，而且是非限制性的，本发明的范围由所附权利要求而不是上述说明限定，因此旨在将落在权利要求的等同要件的含义和范围内的所有变化囊括在本发明内。 
此外，应当理解，虽然本说明书按照实施方式加以描述，但并非每个实施方式仅包含一个独立的技术方案，说明书的这种叙述方式仅仅是为清楚起见，本领域技术人员应当将说明书作为一个整体，各实施例中的技术方案也可以经适当组合，形成本领域技术人员可以理解的其他实施方式。 
序列表
 

Claims (1)

1.一种基于血浆microRNA分析的卵巢癌肿瘤诊断方法，其特征是，包括以下步骤：
(1)建立血浆microRNAs抽提、qRT-PCR定量检测的技术平台；
(2)利用miRNAsmicroarray分析卵巢癌患者和正常志愿者血浆microRNAs表达芯片，寻找差异表达的microRNAs；
(3)利用Northern Blot或qRT-PCR技术，筛选血浆样本中肿瘤特异性高表达的miRNAs作为候选血浆miRNAs标志物；
(4)在卵巢癌患者的血浆中，通过检测血浆中特异高表达的miRNAs含量的变化，结合医学统计学，初步评价与确定了候选血浆miRNAs诊断卵巢癌的能力。
CN201310698673.4A 2013-12-10 2013-12-10 一种基于血浆microRNA分析的卵巢癌肿瘤诊断方法 Pending CN103952465A (zh)

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Cited By (3)

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CN105311633A (zh) * 2015-11-26 2016-02-10 南京市妇幼保健院 一种载microRNA的石墨烯-金纳米复合物及其制备方法和应用
CN110564854A (zh) * 2019-09-09 2019-12-13 晶准生物医药集团有限公司 基于miRNA检测的试剂盒及其在癌症早期筛查中的应用
CN112626209A (zh) * 2020-12-22 2021-04-09 绵竹市人民医院 用于卵巢癌诊断的miRNA标志物、其应用及诊断试剂盒

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CN102292456A (zh) * 2008-12-05 2011-12-21 俄亥俄州立大学研究基金会 用于诊断和治疗卵巢癌的基于microRNA 的方法和组合物

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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105311633A (zh) * 2015-11-26 2016-02-10 南京市妇幼保健院 一种载microRNA的石墨烯-金纳米复合物及其制备方法和应用
CN110564854A (zh) * 2019-09-09 2019-12-13 晶准生物医药集团有限公司 基于miRNA检测的试剂盒及其在癌症早期筛查中的应用
CN110564854B (zh) * 2019-09-09 2023-09-29 晶准生物医药集团有限公司 基于miRNA检测的试剂盒及其在癌症早期筛查中的应用
CN112626209A (zh) * 2020-12-22 2021-04-09 绵竹市人民医院 用于卵巢癌诊断的miRNA标志物、其应用及诊断试剂盒

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