| 猪细环病毒2型环介导等温扩增检测方法的建立 |
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| 引用本文： | 张黎,李凯,田光玲,李德峰,王伟松,何后军.猪细环病毒2型环介导等温扩增检测方法的建立[J].中国畜牧兽医,2015,42(5):1098-1103. |
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| 作者姓名： | 张黎 李凯 田光玲 李德峰 王伟松 何后军 |
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| 作者单位： | 江西农业大学动物科学技术学院, 南昌 330045 |
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| 基金项目： | 2013年度江西省研究生创新专项资金(YC2013-S123) |
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| 摘 要： | 为了能快速、特异的检测猪细环病毒2型(TTSuV2),本研究针对TTSuV2全基因序列的非编码区域和第1个开放性阅读框前端设计了2对引物,建立了TTSuV2的环介导等温扩增(LAMP)检测方法并对反应成分和条件进行了梯度摸索。试验结果显示,该LAMP检测方法最佳反应条件为64 ℃恒温90 min,可特异性检测TTSuV2,与猪细环病毒1型、猪圆环病毒2型、猪瘟病毒、猪繁殖与呼吸综合征病毒和猪博卡病毒无交叉反应,病毒最低检出限为100拷贝/μL。结果表明,建立的LAMP方法具有快速、特异且灵敏的特点,可在TTSuV2快速检测方面提供一定的技术支持。
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| 关 键 词： | 猪细环病毒2型(TTSuV2) 环介导等温扩增 检测 |
| 收稿时间： | 2014-11-17 |
Development of Loop-mediated Isothermal Amplification Assay for Detection of Torque teno sus Virus Type 2 |
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| ZHANG Li,LI Kai,TIAN Guang-ling,LI De-feng,WANG Wei-song,HE Hou-jun.Development of Loop-mediated Isothermal Amplification Assay for Detection of Torque teno sus Virus Type 2[J].China Animal Husbandry & Veterinary Medicine,2015,42(5):1098-1103. |
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| Authors: | ZHANG Li LI Kai TIAN Guang-ling LI De-feng WANG Wei-song HE Hou-jun |
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| Institution: | College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang 330045, China |
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| Abstract: | In order to develop a loop-mediated isothermal amplification (LAMP) assay for rapid and specific detection of Torque teno sus virus type 2 (TTSuV2), two pairs of primers were designed according to the untranslated regions and the part of open reading frame 1 of TTSuV2.The LAMP system was optimized by adjusting the concentrations of some components and reaction conditions.The optimized amplification conditions of LAMP assay was at 64 ℃ for 90 min.The results showed the LAMP assay was specific for TTSuV2 detection, which could achieve a detection limit of 100 copies/μL viral nucleic acid, and no cross-reaction with TTSuV1, PCV2, CSFV, PRRSV and PBoV.In conclusion, this assay was a rapid, specific and sensitive detection technique which could provide a assistance for the rapid detection of TTSuV2. |
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| Keywords: | Torque teno sus virus type 2 (TTSuV2) loop-mediated isothermal amplification (LAMP) detection |
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