CN105734151A - mtDNA拷贝数评估冠状动脉旁路移植术后新发房颤的应用 - Google Patents

mtDNA拷贝数评估冠状动脉旁路移植术后新发房颤的应用 Download PDF

Info

Publication number
CN105734151A
CN105734151A CN201610243957.8A CN201610243957A CN105734151A CN 105734151 A CN105734151 A CN 105734151A CN 201610243957 A CN201610243957 A CN 201610243957A CN 105734151 A CN105734151 A CN 105734151A
Authority
CN
China
Prior art keywords
copy number
coronary artery
artery bypass
atrial fibrillation
dna copy
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CN201610243957.8A
Other languages
English (en)
Inventor
张建
王辉山
徐银丽
徐殊
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Individual
Original Assignee
Individual
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Individual filed Critical Individual
Priority to CN201610243957.8A priority Critical patent/CN105734151A/zh
Publication of CN105734151A publication Critical patent/CN105734151A/zh
Pending legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/118—Prognosis of disease development
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/158—Expression markers

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Wood Science & Technology (AREA)
  • Analytical Chemistry (AREA)
  • Zoology (AREA)
  • Genetics & Genomics (AREA)
  • Engineering & Computer Science (AREA)
  • Pathology (AREA)
  • Immunology (AREA)
  • Microbiology (AREA)
  • Molecular Biology (AREA)
  • Biotechnology (AREA)
  • Biophysics (AREA)
  • Physics & Mathematics (AREA)
  • Biochemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

本发明提供了一种利用外周血线粒体DNA拷贝数评估非体外循环下冠状动脉旁路移植术后新发房颤的方法。该方法步骤为：通过实时定量PCR检测术前冠状动脉旁路移植患者血液中线粒体DNA拷贝数，根据其实施定量PCR结果，可以判断患者术后新发房颤的发生，指导冠状动脉旁路移植术后患者的个体化治疗。

Description

mtDNA拷贝数评估冠状动脉旁路移植术后新发房颤的应用
技术领域
本发明属于医学生物工程技术领域。具体的说，本发明涉及利用循环血中线粒体DNA拷贝数(mtDNAcopynumber)评估非体外循环下冠状动脉旁路移植术后新发房颤的标记应用。
背景技术
术后新发房颤是冠状动脉旁路移植术后最常见的并发症，其发生率在15-50％之间。术后新发房颤与患者长期的病死率关系密切。目前发现，高龄、种族、结构性心肌病、代谢综合症、肥胖等都与术后房颤相关。有研究报道，ROS产生相关酶(NAPDH氧化酶)在术后房颤患者的心房组织中表达明显升高。与ROS产生相关的另一种酶单胺氧化酶(MAO)也被发现在术后房颤患者心房肌中高表达，并且在人的心房肌中的氧化还原平衡中起到重要作用。这说明，氧化应激是在术后房颤患者中更加持久的存在，也暗示，氧化应激与术后房颤关系紧密。(Y.M.Kimet.al2008)
线粒体作为能量产生器官，在细胞的多种功能中发挥着重要作用。有研究发现，线粒体DNA拷贝数的变化与多种疾病相关。一些研究者发现，外周血中线粒体DNA拷贝数与氧化应激的水平相关，并且增加2型糖尿病和心血管疾病的危险。很多研究者也发现，外周血的线粒体DNA拷贝数在结肠癌、乳腺癌和肺癌等多种肿瘤中表达异常。这些均提示，外周血线粒体DNA拷贝数可能作为一个炎症和氧化应激相关的潜在标志物。以上内容均提示外周血线粒体DNA拷贝数可能作为冠状动脉旁路移植术后新发房颤的标志物，预测冠状动脉旁路移植术患者房颤的发生。(J.Mengel-Frometal,,2014)
发明内容
基于上述发现，本发明的首要目的在于提供一种外周血中线粒体DNA拷贝数(mtDNAcopynumber)在评估非体外循环下冠状动脉旁路移植术后新发房颤的标记应用。其目的是解决目前对非体外循环下冠状动脉旁路移植术后新发房颤的生物学诊断的问题。
本发明的技术方案主要包括以下技术方案来实现：
本发明提供了循环血中线粒体DNA拷贝数在评估非体外循环下冠状动脉旁路移植术后新发房颤标记中的应用。
其中，所述的应用是通过实时定量PCR的方法检测冠状动脉旁路移植术患者术前外周血线粒体DNA的拷贝数来实现的。
其中，所述的应用的具体操作步骤为：
1、通过实时定量PCR的方法检测拟行非体外循环冠状动脉旁路移植术患者术前全血中线粒体DNA拷贝数。应用外周血全基因组抽提试剂盒(SangonBiotech)，
引物设计如下：ND1-F5’-CCCTAAAACCCGCCACATCT-3’；ND1-R
5’-GAGCGATGGTGAGAGCTAAGGT-3’，HGB-1-F5’-GTGCACCTGACTCCTGAGGAGA-3’；HGB-1-R5’-CCTTGATACCAACCTGCCCAG-3’.PCR使用FastSYBRGreendye2xAppliedBiosystems，384孔板，10微升反应体系，7900HTFastReal-TimePCR机器。(W.Zhouetal,2014)
2、根据PCR检测得到的非体外循环冠状动脉旁路移植术患者术前全血中线粒体DNA拷贝数，进行分析。方法如下：每个标本重复三个复孔，CT值差值在0.3内，取平均值；外周血线粒体DNA拷贝数2-ΔΔCt(ΔCt＝CtND-1-CtHGB)根据结果，取线粒体DNA拷贝数中位数作为高低之分。
研究结果表明，本发明方法可以提供一种外周血中线粒体DNA拷贝数(mtDNAcopynumber)用于评估非体外循环下冠状动脉旁路移植术后新发房颤的标记应用。
本发明提供了外周血线粒体DNA拷贝数(mtDNAcopynumber)的新用途。实验结果表明：
通过应用实时定量PCR方法检测在外周血中线粒体DNA表达情况发现，在临床样本中，发生术后房颤患者的外周血线粒体DNA拷贝数明显高于非发生房颤患者。
根据实时定量PCR结果，高表达线粒体DNA拷贝数患者术后房颤率发生明显高于低表达线粒体DNA拷贝数患者，表明外周血中线粒体DNA拷贝数与冠状动脉旁路移植术后患者明显相关。
因此，外周血中线粒体DNA拷贝数是判断非体外循环冠状动脉旁路移植术后新发房颤的独立危险因素。
总之，根据实时定量PCR结果，外周血线粒体DNA拷贝数是一个可以预测非体外循环冠状动脉旁路移植术后新发房颤的标志物。
另外，本发明还提供了一种可以评估非体外循环冠状动脉旁路移植术后新发房颤的方法，可以指导患者进行冠状动脉旁路移植术后预防房颤发生的方法。
本发明为外周血线粒体DNA拷贝数PCR试剂盒的临床应用提供了新的思路。
参考文献
Y.M.Kim,H.Kattach,C.Ratnatunga,R.Pillai,K.M.Channon,B.Casadei,Associationofatrialnicotinamideadeninedinucleotidephosphateoxidaseactivitywiththedevelopmentofatrialfibrillationaftercardiacsurgery,JAmCollCardiol,51(2008)68-74.
J.Mengel-From,M.Thinggaard,C.Dalgard,K.O.Kyvik,K.Christensen,L.Christiansen,MitochondrialDNAcopynumberinperipheralbloodcellsdeclineswithageandisassociatedwithgeneralhealthamongelderly,HumGenet,133(2014)1149-1159.
W.Zhou,M.Zhu,M.Gui,L.Huang,Z.Long,L.Wang,H.Chen,Y.Yin,X.Jiang,Y.Dai,Y.Tang,L.He,K.Zhong,PeripheralbloodmitochondrialDNAcopynumberisassociatedwithprostatecancerriskandtumorburden,PLoSOne,9(2014)e109470.
附图说明
图1是冠状动脉旁路移植术前患者外周血中线粒体DNA拷贝数的表达
图1是利用实时定量PCR方法，检测在非体外循环下冠状动脉旁路移植术后发生房颤组(80例)和未发生房颤组(255例)术前外周血线粒体DNA的表达情况。
具体实施方式
现结合实施例和附图，对本发明作进一步描述，但本发明的实施并不仅限于此。
实施例1线粒体DNA拷贝数在非体外循环冠状动脉旁路移植术患者外周血的表达
在沈阳军区总医院心血管外科选取2015年1月份至2015年6月份行非体外循环冠状动脉旁路移植术的患者335名，收取术前外周血3毫升，使用全血基因组抽提试剂盒(SangonBiotech)，对抽提的基因组进行实时定量PCR检测基因组中线粒体DNA的拷贝数表达。
具体方法如下：通过实时定量PCR的方法检测拟行非体外循环冠状动脉旁路移植术患者术前全血中线粒体DNA拷贝数。应用外周血全基因组抽提试剂盒(SangonBiotech)，引物设计如下：ND1-F5’-CCCTAAAACCCGCCACATCT-3’；ND1-R
5’-GAGCGATGGTGAGAGCTAAGGT-3’，HGB-1-F5’-GTGCACCTGACTCCTGAGGAGA-3’；HGB-1-R5’-CCTTGATACCAACCTGCCCAG-3’.PCR使用FastSYBRGreendye2xAppliedBiosystems，384孔板，10微升反应体系，7900HTFastReal-TimePCR机器。(W.Zhouetal,2014)
根据PCR检测得到的非体外循环冠状动脉旁路移植术患者术前全血中线粒体DNA拷贝数，进行分析。方法如下：每个标本重复三个复孔，CT值差值在0.3内，取平均值；外周血线粒体DNA拷贝数2-ΔΔCt(ΔCt＝CtND-1-CtHGB)根据结果，取线粒体DNA拷贝数中位(17.16)数作为高低之分。
每个标本重复三个复孔，CT值差值在0.3内，取平均值；外周血线粒体DNA拷贝数2-ΔΔCt(ΔCt＝CtND-1-CtHGB).根据结果，取线粒体DNA拷贝数中位数(17.16)作为高低之分。
研究显示，在术后发生房颤的患者中线粒体DNA拷贝数程明显高表达。这表明，应用外周血线粒体DNA拷贝数可以区分非体外循环下冠状动脉旁路移植术后患者发生房颤与否，具有良好的特异性和敏感性。
实施例2卡方检验分析线粒体DNA拷贝数与临床资料相关性
在实施例1的患者中，分析线粒体DNA的拷贝数表达，取Ct值中位数17.16为分界点，高于17.16的为线粒体DNA拷贝数增加，低于17.16的为线粒体DNA拷贝数降低。
表1线粒体DNA拷贝数与临床资料相关性分析
结果显示，非体外循环冠状动脉旁路移植术前患者线粒体DNA拷贝数与患者年龄和术后房颤发生明显相关。
实施例3Logistic回归分析影响非体外循环冠状动脉旁路移植术后发生房颤的因素
在实施例1的患者中，应用SPSS18.0分析软件进行二元Logistic回归分析。
表2.单因素和多因素Logistic回归分析
结果显示，患者术前外周血线粒体DNA拷贝数是影响非体外循环冠状动脉旁路移植术后发生房颤的独立危险因素。

Claims (3)

1.循环血中线粒体DNA拷贝数在评估非体外循环下冠状动脉旁路移植术后新发房颤标记中的应用。
2.如权利要求1所述的循环血中线粒体DNA拷贝数在评估非体外循环下冠状动脉旁路移植术后新发房颤标记中的应用，其特征在于，所述的应用是通过实时定量PCR的方法检测冠状动脉旁路移植术患者术前外周血线粒体DNA的拷贝数来实现的。
3.如权利要求1或2所述的循环血中线粒体DNA拷贝数在评估非体外循环下冠状动脉旁路移植术后新发房颤标记中的应用，其特征在于，所述应用的具体操作步骤为：
(1)通过实时定量PCR的方法检测非体外循环冠状动脉旁路移植术患者术前全血中线粒体DNA拷贝数；
(2)根据PCR检测得到的非体外循环冠状动脉旁路移植术患者术前全血中线粒体DNA拷贝数，进行分析；方法如下：每个标本重复三个复孔，CT值差值在0.3内，取平均值；根据结果，取线粒体DNA中位数作为高低之分；高线粒体DNA拷贝数组，冠状动脉旁路移植术后患者易发生房颤；低线粒体DNA拷贝数组，冠状动脉旁路移植术后患者不易发生房颤。
CN201610243957.8A 2016-04-19 2016-04-19 mtDNA拷贝数评估冠状动脉旁路移植术后新发房颤的应用 Pending CN105734151A (zh)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201610243957.8A CN105734151A (zh) 2016-04-19 2016-04-19 mtDNA拷贝数评估冠状动脉旁路移植术后新发房颤的应用

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201610243957.8A CN105734151A (zh) 2016-04-19 2016-04-19 mtDNA拷贝数评估冠状动脉旁路移植术后新发房颤的应用

Publications (1)

Publication Number Publication Date
CN105734151A true CN105734151A (zh) 2016-07-06

Family

ID=56254587

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201610243957.8A Pending CN105734151A (zh) 2016-04-19 2016-04-19 mtDNA拷贝数评估冠状动脉旁路移植术后新发房颤的应用

Country Status (1)

Country Link
CN (1) CN105734151A (zh)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2020188938A1 (ja) * 2019-03-20 2020-09-24 国立大学法人 東京医科歯科大学 頻脈性不整脈検査方法及び頻脈性不整脈検査システム

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102480951A (zh) * 2009-04-17 2012-05-30 卡迪罗治疗公司 用于治疗局部缺血病症和与线粒体功能相关病症的方法和组合物

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102480951A (zh) * 2009-04-17 2012-05-30 卡迪罗治疗公司 用于治疗局部缺血病症和与线粒体功能相关病症的方法和组合物

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
M.ZAKKAR等: "inflammation, oxidative stress and postoperative atrial fibrillation in cardiac surgery", 《PHARMACOLOGY AND THERAPEUTIC》 *
PO-HAN LIN等: "OXIDATIVE DAMAGE TO MITOCHONDRIAL DNA IN ATRIAL MUSCLE OF PATIENTS WITH ATRIAL FIBRILLATION", 《FREE RADICAL BIOLOGY & MEDICINE》 *
WEIMIN ZHOU等: "Peripheral Blood Mitochondrial DNA Copy Number is Association with Prostate Cancer Risk and Tumor Burden", 《PLOS ONE》 *
张芃等: "心房肌线粒体功能异常增加冠脉旁路移植术后新发房颤的风险", 《中国分子心脏病学》 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2020188938A1 (ja) * 2019-03-20 2020-09-24 国立大学法人 東京医科歯科大学 頻脈性不整脈検査方法及び頻脈性不整脈検査システム

Similar Documents

Publication Publication Date Title
McManus et al. Plasma microRNAs are associated with atrial fibrillation and change after catheter ablation (the miRhythm study)
Pedrotty et al. Transcriptomic biomarkers of cardiovascular disease
Junejo et al. Cardiopulmonary exercise testing for preoperative risk assessment before pancreaticoduodenectomy for cancer
Christman et al. Yield of downstream tests after exercise treadmill testing: a prospective cohort study
CN105002286A (zh) 与高血压和/或心血管疾病发病风险相关的多个单核苷酸多态性位点及相关应用
Alonso et al. Serum metabolomics and incidence of atrial fibrillation (from the Atherosclerosis Risk in Communities Study)
Darling et al. Survival after hospital discharge for ST-segment elevation and non-ST-segment elevation acute myocardial infarction: a population-based study
Filardo et al. Underestimation of the incidence of new-onset post–coronary artery bypass grafting atrial fibrillation and its impact on 30-day mortality
Sheng et al. Correlation between fragmented QRS and the short-term prognosis of patients with acute myocardial infarction
Tse et al. Meta-analysis of Tpeak–Tend and Tpeak–Tend/QT ratio for risk stratification in congenital long QT syndrome
Vrba et al. DNA methylation biomarkers discovered in silico detect cancer in liquid biopsies from non-small cell lung cancer patients
Engin et al. Visceral adiposity index and prognostic nutritional index in predicting atrial fibrillation after on-pump coronary artery bypass operations: a prospective study
Bellam et al. Prognostic significance of impaired chronotropic response to pharmacologic stress Rb-82 PET
Zhang et al. Prognostic significance of regulatory T lymphocytes in patients with hepatocellular carcinoma
Erkus et al. The association between plasma thiol levels and left ventricular diastolic dysfunction in patient with hypertension
Snelder et al. Cross-sectional and prospective follow-up study to detect early signs of cardiac dysfunction in obesity: protocol of the CARDIOBESE study
Yoon et al. Prognostic value of baseline sarcopenia on 1-year mortality in patients undergoing transcatheter aortic valve implantation
CN105734151A (zh) mtDNA拷贝数评估冠状动脉旁路移植术后新发房颤的应用
Heisterberg et al. Multiple blood samples in elite soccer players. Is it worthwhile?
CN107653312A (zh) 与血脂水平和冠心病相关的rs7901016检测系统及相关应用
Eaton et al. Recall processes for biliary cytology in primary sclerosing cholangitis
Li et al. Electrocardiography is useful to predict postoperative ventricular arrhythmia in patients undergoing cardiac surgery: a retrospective study
CN107653311B (zh) 与血脂水平相关的单核苷酸多态性rs4883263检测系统及相关应用
Cuspidi et al. Electrocardiographic diagnosis of left-ventricular hypertrophy: good news for the clinician?
Sun et al. Amino acid profiling as a screening and prognostic biomarker in active tuberculosis patients

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
WD01 Invention patent application deemed withdrawn after publication
WD01 Invention patent application deemed withdrawn after publication

Application publication date: 20160706