改良多重实时荧光定量PCR检测流感A型和B型病毒 |
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引用本文: | 邸红芹,杨永辉,朱桂云,王晓玲,杨庆志,范海霞,王亮,张辉.改良多重实时荧光定量PCR检测流感A型和B型病毒[J].河北医药,2016(23):3529-3532. |
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作者姓名: | 邸红芹 杨永辉 朱桂云 王晓玲 杨庆志 范海霞 王亮 张辉 |
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作者单位: | 1. 河北省胸科医院, 石家庄市,050041;2. 河北省唐山市丰润区第二人民医院;3. 河北省三河市中医医院 |
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基金项目: | 河北省医学科学研究重点课题(编号20150147) |
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摘 要: | 目的:建立改良多重实时荧光定量PCR体系快速检测流感A型和B型病毒。方法根据生物信息学分析结果选定流感A型和B型病毒的靶序列,建立MRT-PCR检测体系。构建质粒标准品,评估所建立体系的灵敏度、特异性和重复性。结果成功建立了基于同源加尾系统和Taqman-分子灯标探针的改良多重实时荧光定量PCR检测体系;该方法最低检测限为102 copies/μl,特异性良好,重复性良好,变异系数(CV)为0.99%~2.50%。结论建立了快速、敏感、特异、稳定地改良多重实时荧光定量PCR( MRT-PCR)检测体系,具有良好的临床应用前景。
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关 键 词: | 多重实时荧光定量PCR 同源加尾系统 Taqman-分子灯标 流感病毒 |
Detection of influenza virus A and B by modified multiplex real-time fluorescence PCR assay |
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Abstract: | Objective To set up a modified multiplex real-time fluorescence quantitative PCR ( MRT-PCR) assay to detect influenza virus A and B rapidly .Methods The MRT-PCR assay was established according to target sequence of influenza virus A and B specific gene selected by bioinformatics analysis results .The standard products and quality control samples were built to evaluate the sensitivity , specificity and repeatability of the system .Results The MRT-PCR detection system was successfully established based on Taqman-molecular beacon ( Taqman-MB) probe and Homo-Taq Assidted Non-Dimer ( HAND) system.The detection limit of the assay was 102 copies/μl,which showed good specificity and repeatability , with coefficient of variation (CV) being 0.99%~2.50%.Conclusion A fast,specific,sensitive,stable, modified MRT-PCR detection system is successfully established ,which possesses favourable application prospect in clinical practice . |
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Keywords: | MRT-PCR HAND system Taqman-molecular beacon influenza virus |
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