| 实时荧光定量PCR检测乙肝病毒DNA方法在绒促性素和尿促性素生产中的应用 |
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| 引用本文： | 吴利红,王灿,邵泓,陈钢.实时荧光定量PCR检测乙肝病毒DNA方法在绒促性素和尿促性素生产中的应用[J].药物分析杂志,2011(10). |
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| 作者姓名： | 吴利红 王灿 邵泓 陈钢 |
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| 作者单位： | 上海市食品药品检验所; |
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| 摘 要： | 目的:验证实时荧光定量PCR(Real-time PCR)检测绒促性素(HCG)和尿促性素(HMG)生产工艺中尿液、粗品和原料药中乙肝病毒DNA(HBV DNA)的方法,并用该方法对HCG和HMG生产工艺中关键点进行控制。方法:在不同浓度的乙肝阳性血中添加HCG或HMG的尿液、粗品和原料药,用Real-time PCR方法检测添加前后不同浓度乙肝阳性血中HBV DNA的浓度,计算出HBV DNA回收率,同时验证该方法的线性、重复性和灵敏度;用Real-time PCR方法对尿液、粗品和原料药样品进行HBV DNA检测,将检测出阳性的样品用ELISA方法检测,比较2种方法的检测结果差异。结果:Real-time PCR检测HBV DNA在104～107IU.mL-1范围内线性关系良好,r2=0.9969;HCG或HMG尿液、粗品和原料药中添加不同浓度乙肝阳性血,HBV DNA回收率为94.9%～103.8%;3次重复实验相对标准偏差(RSD)均小于5%;最低检出限为500 IU.mL-1。Real-time PCR检测出阳性的样品用ELISA方法检测乙肝表面抗原(HBsAg)均为阴性。结论:Real-time ...
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| 关 键 词： | 实时荧光定量PCR 乙肝病毒DNA 绒促性素 尿促性素 |
Application of Real-time fluorescence PCR detecting HBV DNA during the production flow of HCG and HMG |
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| WU Li-hong,WANG Can,SHAO Hong,CHEN Gang.Application of Real-time fluorescence PCR detecting HBV DNA during the production flow of HCG and HMG[J].Chinese Journal of Pharmaceutical Analysis,2011(10). |
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| Authors: | WU Li-hong WANG Can SHAO Hong CHEN Gang |
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| Institution: | WU Li-hong,WANG Can,SHAO Hong,CHEN Gang(Shanghai Institute for Food and Drug Control,Shanghai 201203,China) |
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| Abstract: | Objective:To verify the method of Real-time fluorescence PCR to detect HBV DNA during the production flow of HCG and HMG.Methods:HCG or HMG was added into positive blood containing different concentration HBV,and the Real-time fluorescence PCR was used to detect the concentration of HBV DNA.The recovery was calculated,and the linear range,repetition and sensitivity of the method were verified.The positive results detected by Real-time PCR would be detected by ELISA again,and the results of two methods would... |
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| Keywords: | real-time fluorescence PCR HBV HCG HMG |
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