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| 鸡传染性支气管炎病毒荧光定量PCR标准曲线的建立 | |
| 引用本文： | 武聚富,张中文,毛东有,沈红,陆彦,况玲,吴国娟.鸡传染性支气管炎病毒荧光定量PCR标准曲线的建立[J].北京农学院学报,2009,24(2):40-43. |
| 作者姓名： | 武聚富 张中文 毛东有 沈红 陆彦 况玲 吴国娟 |
| 作者单位： | 1. 新疆农业大学乌鲁木齐,830052;北京农学院动物科学技术系,北京,102206 2. 北京农学院动物科学技术系,北京,102206 3. 新疆农业大学乌鲁木齐,830052 |
| 基金项目： | 北京市自然科学基金，国家自然科学基金，北京市属高校人才强教计划 |
| 摘 要： | 笔者针对传染性支气管炎病毒（IBV）M41株中编码核衣壳蛋白的N基因设计并合成一对引物，通过构建重组阳性标准质粒的方法，成功建立SYBR GreenⅠ荧光定量PCR方法检测IBV的标准曲线。该方法所建立的曲线可检测到初始模板中5×10^2copies／μL的病毒核酸，与常规PCR相比敏感性大大提高。笔者建立的荧光定量PCR检测方法具有灵敏度高、特异性强、重复性好等特点，可用于传染性支气管炎的临床诊断和健康带毒鸡群的检测。 |
| 关 键 词： | 传染性支气管炎病毒 荧光定量PCR 病毒检测 标准曲线 |
Construction of Standard Curve for Real-time PCRAssay of Infectious Bronchitis Virus |
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| WU Ju-fu,ZHANG Zhong-wen,MAO Dong-you,SHEN Hong,LU Yan,KUANG Ling,WU Guo-juan.Construction of Standard Curve for Real-time PCRAssay of Infectious Bronchitis Virus[J].Journal of Beijing Agricultural College,2009,24(2):40-43. | |
| Authors: | WU Ju-fu ZHANG Zhong-wen MAO Dong-you SHEN Hong LU Yan KUANG Ling WU Guo-juan |
| Institution: | 1.Xinjiang Agricultural University;Urumqi 830052;China;2.Department of Animal Science and Veterinary Medicine;Beijing University of Agriculture;Beijing 102206;China |
| Abstract: | In this study,the authors constructed standard curve of Real-time PCR assay for detecting infectious bronchitis virus(IBV).In this experiment,the dye of SYBR GreenⅠ was used.A pair of primers was designed according to the sequences of the N gene which coding nucleocapsid protein of M41strain.And authors used positive recombinant plasmid as standard to construct the standard curve.This method had a detection limit of 5×102 copies/μL viral RNA,its sensitivity greatly enhanced compared to conventional PCR.The results of the test showed that the Real-time PCR could be used for the clinical diagnosis of IBV infection because of its high sensitivity,specificity and repeatability. |
| Keywords: | infectious bronchitis virus(IBV) real-time PCR virus detection standard curve |
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