Development of Real-Time PCR Assays for the Quantitative Detection of CD81 Receptor Gene of Hepatitis C Virus in Tupaia Belangeri

2009 2nd International Conference on Biomedical Engineering and Informatics Pub Date : 2009-10-30 DOI:10.1109/BMEI.2009.5305205

Yue Feng, Yue Feng, J. Bai, X. Xia, Wenhua Zhao, Jiejie Dai, Xiaomei Sun, Qihan Li

{"title":"Development of Real-Time PCR Assays for the Quantitative Detection of CD81 Receptor Gene of Hepatitis C Virus in Tupaia Belangeri","authors":"Yue Feng, Yue Feng, J. Bai, X. Xia, Wenhua Zhao, Jiejie Dai, Xiaomei Sun, Qihan Li","doi":"10.1109/BMEI.2009.5305205","DOIUrl":null,"url":null,"abstract":"—A real-time PCR assay based on the TaqMan chemistry was developed for reliable and quantitative detection of CD81 in Tupaia belangeri. The assay is performed with the ABI 7300 system using TaqMan probe and primers amplifying 118bp CD81 fragment of Tupaia belangeri. The standard curve for quantitation of target gene showed linearity over an at least 5-log DNA concentration range, represents10 3 to10 7 copies per reaction, with a correlation coefficient of 0.9996 (the slopes value-3.39 VS-3.32). Moreover, this protocol enabled detection of as little as 10 copies of CD81 cDNA in Tupaia belangeri liver tissues. The overall % coefficient of variation (%CV) for this assay was lower 5% with statistical significance due to in intra-assay 1.08% and inter-assay 0.16%, which indicated its high reproducibility. The new assay greatly improves current detection methods for CD81 evaluation, and this is the first report on the standardization and evaluation of a Tupaia belangeri CD81 RNA quantitation assay from China. As TaqMan real-time PCR enable the rapid and easy processing of a large number of samples, and can be used as a tool for monitoring progression of CD81 expression.","PeriodicalId":6389,"journal":{"name":"2009 2nd International Conference on Biomedical Engineering and Informatics","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"2009-10-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"2009 2nd International Conference on Biomedical Engineering and Informatics","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1109/BMEI.2009.5305205","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}

引用次数: 0

Abstract

—A real-time PCR assay based on the TaqMan chemistry was developed for reliable and quantitative detection of CD81 in Tupaia belangeri. The assay is performed with the ABI 7300 system using TaqMan probe and primers amplifying 118bp CD81 fragment of Tupaia belangeri. The standard curve for quantitation of target gene showed linearity over an at least 5-log DNA concentration range, represents10 3 to10 7 copies per reaction, with a correlation coefficient of 0.9996 (the slopes value-3.39 VS-3.32). Moreover, this protocol enabled detection of as little as 10 copies of CD81 cDNA in Tupaia belangeri liver tissues. The overall % coefficient of variation (%CV) for this assay was lower 5% with statistical significance due to in intra-assay 1.08% and inter-assay 0.16%, which indicated its high reproducibility. The new assay greatly improves current detection methods for CD81 evaluation, and this is the first report on the standardization and evaluation of a Tupaia belangeri CD81 RNA quantitation assay from China. As TaqMan real-time PCR enable the rapid and easy processing of a large number of samples, and can be used as a tool for monitoring progression of CD81 expression.

查看原文本刊更多论文

丙型肝炎病毒CD81受体基因实时荧光定量检测方法的建立

-建立了基于TaqMan化学的实时荧光定量PCR方法，可靠、定量地检测了图帕亚belangeri中CD81的含量。采用ABI 7300系统，使用TaqMan探针和引物扩增图皮亚belangeri的118bp CD81片段。目的基因定量标准曲线在至少5对数DNA浓度范围内呈线性关系，即每次反应为103 ~ 10.7个拷贝，相关系数为0.9996(斜率为3.39 vs . 3.32)。此外，该方案能够在图帕亚贝兰格里肝组织中检测到10个CD81 cDNA拷贝。结果表明，该方法的总变异系数(%CV)比原方法低5%，组内变异系数为1.08%，组间变异系数为0.16%，具有统计学意义，重复性高。新方法大大改进了现有的CD81评价检测方法，这是中国首个关于图帕亚贝兰杰CD81 RNA定量分析的标准化和评价报告。由于TaqMan实时PCR能够快速简便地处理大量样品，可以作为监测CD81表达进展的工具。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文求助全文

来源期刊

2009 2nd International Conference on Biomedical Engineering and Informatics

自引率

0.00%

发文量