| 锦鲤疱疹病毒荧光定量PCR检测方法的建立 |
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| 引用本文： | 张艳,孟庆峰,钱爱东,王伟利.锦鲤疱疹病毒荧光定量PCR检测方法的建立[J].吉林畜牧兽医,2010,31(4):11-14. |
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| 作者姓名： | 张艳 孟庆峰 钱爱东 王伟利 |
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| 作者单位： | 1. 吉林农业大学动物科技学院,吉林长春,130118
2. 吉林出入境检验检疫局,吉林长春,130062 |
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| 基金项目： | 吉林省科技厅项目(20080218) |
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| 摘 要： | 根据锦鲤疱疹病毒(KHV)胸苷激酶(TK)基因的保守序列设计一对引物和相应的TaqMan探针,建立荧光定量PCR方法,用于锦鲤疱疹病毒快速、灵敏、可定量的检测手段。构建并制备实时荧光定量PCR的标准品,对反应体系进行优化,并做特异性,敏感性和重复性试验。结果表明,成功构建荧光定量PCR标准品,建立荧光定量标准曲线,标准曲线的相关系数为0.992,所建立的荧光定量PCR方法特异性强、敏感性高,重复性好。
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| 关 键 词： | 锦鲤疱疹病毒 荧光定量PCR |
Establishment of a Flurogenic Quantitative Polymerase Chain Reaction Assay for Detection of Kio Hepesvirus |
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| ZHANG Yan,MENG Qing-feng,QIAN Ai-dong,WANG Wei-li.Establishment of a Flurogenic Quantitative Polymerase Chain Reaction Assay for Detection of Kio Hepesvirus[J].Jilin Animal Science and Veterinary Medicine,2010,31(4):11-14. |
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| Authors: | ZHANG Yan MENG Qing-feng QIAN Ai-dong WANG Wei-li |
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| Institution: | 1.Jilin Agricutural University;College of Animal Science and Technology;Changchun Jilin 130118;2. Jilin Exit&Entry Inspection and Quarantine Bureau;Changchun Jilin 130062 |
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| Abstract: | A set of primers and TaqMan probe for Fluorescent Quantitation PCR were designed to detect the conservative sequence of Kio Hepesvirus thymidine kinase gene. The standard recombinant plasmids for TK gene were constructed as reference standard used in absolute quantification assay. The reaction conditions of FQ-PCR were optimized. The quantitative curve was established using the reference standard plasmids. The FQ-PCR method was tested for sensitivity,repeatability and specificity by amplification the DNA of... |
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| Keywords: | Koi Herpesvirus Flurogenic Quantitative Polymerase Chain Reaction |
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