| Real-time PCR检测核酸疫苗中宿主基因组残留DNA |
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| 引用本文： | 李亮助,刘勇,王贻杰,程海,孙茂盛,邵一鸣.Real-time PCR检测核酸疫苗中宿主基因组残留DNA[J].粉末涂料与涂装,2007,20(6):439-441,446. |
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| 作者姓名： | 李亮助 刘勇 王贻杰 程海 孙茂盛 邵一鸣 |
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| 作者单位： | 中国医学科学院中国协和医科大学医学生物学研究所分子生物学研究室 (昆明650118)中国疾病预防控制中心性病艾滋病预防控制中心传染病预防控制国家重点实验室(北京100050)(李亮助)，中国疾病预防控制中心性病艾滋病预防控制中心传染病预防控制国家重点实验室 (北京100050)(刘勇，王贻杰，程海，邵一鸣)，中国医学科学院中国协和医科大学医学生物学研究所分子生物学研究室 (昆明650118)(孙茂盛) |
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| 摘 要： | 目的建立Real-time PCR方法,用于定量检测核酸疫苗中宿主基因组的残留DNA。方法以Lightcycler平台为基础,选择大肠杆菌23S核糖体RNA基因为靶标基因设计扩增引物,建立基于SYBR GreenⅠ荧光染料的Real-time PCR检测方法,并用于核酸疫苗纯化过程中间产物的检测。结果整个检测过程可在30min内完成,特异性强,检测灵敏度可达10fg/μl,其标准曲线的相关系数为-0.99。结论该方法可用于核酸疫苗中宿主基因组残留DNA的检测。
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| 关 键 词： | Real-time PCR 宿主菌基因组DNA 核酸疫苗 |
| 文章编号： | 1004-5503（2007）06-439-04 |
| 收稿时间： | 2006-09-11 |
| 修稿时间： | 2006-09-11 |
Determination of Residual Host Genomic DNA in DNA Vaccine by Real-time PCR |
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| LI Liang-zhu, LIU Yong, WANG Yi-jie, et al.Determination of Residual Host Genomic DNA in DNA Vaccine by Real-time PCR[J].Chinese Journal of Biologicals,2007,20(6):439-441,446. |
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| Authors: | LI Liang-zhu LIU Yong WANG Yi-jie |
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| Abstract: | Objective To develop a real-time PCR for determination of residual host genomic DNA in DNA vaccine.MethodsAmplify residual host genomic DNA from DNA vaccine by PCR using designed primers target to E.coli 23S ribosome RNA gene and determine the real-time fluorescence intensity of amplified product.Determine the intermediate product during purification of DNA vaccine by the developed real-time PCR.Results The whole determination was completed within 30 min.The developed real-time PCR showed good specificity and a sensitivity of 10 fg/μl.The correlation coefficient of standard curve of the method was-0.99.Conclusion The developed real-time PCR may be used for the determination of residual host genomic DNA in DNA vaccine. |
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| Keywords: | Real-time PCR |
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