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| H9亚型禽流感病毒荧光定量RT-PCR检测方法的建立 | |
| 引用本文： | 陆桂平,曹军平,王涛,赵明军,赵国,刘秀梵. H9亚型禽流感病毒荧光定量RT-PCR检测方法的建立[J]. 畜牧与兽医, 2011, 43(7) |
| 作者姓名： | 陆桂平 曹军平 王涛 赵明军 赵国 刘秀梵 |
| 作者单位： | 1. 江苏畜牧兽医职业技术学院,江苏泰州,225300 2. 江苏畜牧兽医职业技术学院,江苏泰州225300;扬州大学兽医学院,江苏扬州225009 3. 扬州大学兽医学院,江苏扬州,225009 |
| 基金项目： | 国家自然科学基金重点项目(30630048) |
| 摘 要： | 根据H9亚型禽流感病毒HA基因上的保守序列,设计合成引物,以阳性H9亚型禽流感病毒HA基因重组质粒为标准品做标准曲线,建立了荧光定量逆转录聚合酶链反应检测方法。结果表明:本试验建立的标准曲线循环阈值(Ct值)与模板浓度具有良好的线性关系,相关系数为0.997,灵敏度约为6拷贝/μL,对新城疫病毒和其他禽病病毒无交叉反应,特异性好,重复性佳,对193份临床泄殖腔棉拭样品的检测,其结果与经典病毒分离方法符合率大于90.0%。该方法为H9亚型禽流感病毒检测提供了一种特异、敏感、快速、较便宜、高通量、生物安全性好的定量检测手段,将在禽流感病毒临床样品快速筛检、流行病学监测等方面显示良好的应用前景。 |
| 关 键 词： | 禽流感病毒 H9亚型 荧光定量RT-PCR |
Development of a Real-time RT-PCR assay for detection of avian influenza virus H9 subtype |
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| LU Gui-ping,CAO Jun-ping,WANG Tao,ZHAO Ming-jun,ZHAO Guo,LIU Xiu-fan. Development of a Real-time RT-PCR assay for detection of avian influenza virus H9 subtype[J]. Animal Husbandry & Veterinary Medicine, 2011, 43(7) | |
| Authors: | LU Gui-ping CAO Jun-ping WANG Tao ZHAO Ming-jun ZHAO Guo LIU Xiu-fan |
| Affiliation: | LU Gui-ping1,CAO Jun-ping1,2,WANG Tao1,ZHAO Ming-jun2,ZHAO Guo2,LIU Xiu-fan2(1.Jiangsu Vocational and Technical College of Animal Husbandry and Veterinary,Taizhou 225300,China,2.School of Veterinary Medicine,Yangzhou University,Yangzhou 225009,China) |
| Abstract: | According to the conservative region of HA gene of H9 subtype avian influenza virus(AIV),a pair of primers was designed and synthesized.A serial of ten fold dilutions of positive plasmid was prepared and used as standard.The standard curve revealed the good linear relationship between Ct(cycle threshold) and template concentration(R2=0.997).No cross-reaction was detected against other avian viruses.The detection limit was 6 copies of AIV genome.The coincidence rate was more than 90.0% with the traditional v... |
| Keywords: | avian influenza virus(AIV) H9 subtype Real-time RT-PCR |
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