| 肝损伤相关循环miRNAs检测方法的建立与初步应用 |
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| 引用本文： | 王丹妮,黄世峰,府伟灵,张莉萍. 肝损伤相关循环miRNAs检测方法的建立与初步应用[J]. 中华医院感染学杂志, 2011, 21(10): 1984-1987 |
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| 作者姓名： | 王丹妮 黄世峰 府伟灵 张莉萍 |
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| 作者单位： | 1. 重庆医科大学附属第一医院检验科,重庆,400016
2. 第三军医大学第一附属医院检验科,重庆,400038 |
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| 基金项目： | 国家自然科学基金，重庆市自然科学基金，重庆医科大学附属第一医院院内医学科学基金 |
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| 摘 要： | 目的建立可靠的循环mi RNA定量技术,并探讨在检测肝损伤相关mi RNA中的应用价值。方法常规收集正常人血清(浆)标本,mirVanaPARIS试剂盒法抽提血清(浆)小RNA,以mi RNA特异引物引导逆转录,通过Taq Man实时荧光定量PCR对内参U6及mi RNAs进行检测。结果常规收集的临床正常血清标本中U6、mi R-122、mi R-192均能实现特异扩增及定量,相应的平均Ct值约分别为33、27和30,对3份不同留置时间人血清标本定量PCR结果显示,mi R-122、mi R-192和U6的丰度相对稳定;临床肝损伤患者循环mi R-122批内和批间差异分别为1.5%、2.5%,用同法检测胆汁淤积肝损伤模型小鼠循环mi R-122结果显示,循环mi R-122在造模后1 d和3 d增高明显。结论实时定量PCR技术为研究mi RNA提供了较好的技术平台。
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| 关 键 词： | 循环miRNA 实时荧光定量PCR 肝损伤 |
Establishment and initial application of circulating miRNAs quantitive technique in detecting liver injury |
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| WANG Dan-ni,HUANG Shi-feng,FU Wei-ling,ZHANG Li-ping. Establishment and initial application of circulating miRNAs quantitive technique in detecting liver injury[J]. Chinese Journal of Nosocomiology, 2011, 21(10): 1984-1987 |
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| Authors: | WANG Dan-ni HUANG Shi-feng FU Wei-ling ZHANG Li-ping |
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| Affiliation: | WANG Dan-ni,HUANG Shi-feng,FU Wei-ling,ZHANG Li-ping(The First Affiliated Hospital of Chongqing Medical University,Chongqing 400016,China) |
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| Abstract: | OBJECTIVE To establish a reliable platform for the quantification of circulating miRNA,and explore its application value in the detection of circulating miRNA during liver injury.METHODS Normal serum(plasma) samples were collected,and small RNA was extracted using the mirVana PARIS kit.U6 and target miRNAs were reverse translated by specific primers and real-time fluorescence quantitative polymerase chain reaction was adopted to quantify U6 and target miRNAs.RESULTS U6,miR-122 and miR-192 were amplified and... |
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| Keywords: | Circulating miRNA Real-time fluorescence quantitative PCR Liver injury |
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