孔德英,孙涛,李应国,滕少娜,陆丽华,邓朝晖.柑橘黄龙病LAMP快速检测方法的建立[J].中国南方果树,2013,42(1): |
柑橘黄龙病LAMP快速检测方法的建立 |
Development and Evaluation of A Loop-Mediated Isothermal Amplification Method for the Rapid Detection of Huanglongbing |
投稿时间:2012-11-21 修订日期:2012-12-04 |
DOI: |
中文关键词: 柑橘黄龙病 环介导等温扩增 检测 |
英文关键词:Huanglongbing loop-mediated isothermal amplification detect |
基金项目:国家质检总局科研项目 |
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中文摘要: |
本文根据GenBank中柑橘黄龙病菌的外膜蛋白(omp)基因序列,设计了1套环介导等温扩增引物,建立了柑橘黄龙病的环介导等温扩增检测方法。该方法敏感度可达25pg/μL,与实时荧光PCR方法灵敏度相同。全部反应只需一个可控温的水浴锅在60min内即可完成,通过肉眼观察颜色即可直接判定结果。本研究建立的环介导等温扩增方法简便、快速、灵敏、特异,适用于基层工作站的柑橘黄龙病鉴定工作。 |
英文摘要: |
Based on the conserved sequences of the omp gene of Candidatus Liberobacter asiaticum,a set of the loop-mediated isothermal amplification(LAMP) primers was designed to amplify the target genes.The LAMP assay was established to detect HLB.The results showed that the detection limit of HLB-LAMP was 25pg/μL, was the same as real-time PCR method. The amplification could be finished by water bath within 60 minitues,and the result could be judged visualized by naked eyes.These results suggested that the HLB-LAMP method was simple,rapid,sensitive,specific,and suitable for primary workstation to detect HLB. |
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