| 吴 薇1, 刘 斐2, 李开国1, 刘 欢1*.樱桃病毒A陕西分离物基因组扩增及RT-LAMP检测体系的建立[J].植物保护,2026,(1):258-265. |
| 樱桃病毒A陕西分离物基因组扩增及RT-LAMP检测体系的建立 |
| Genome amplification and establishment of an RT-LAMP detection system for cherry virus A isolate from Shaanxi |
| 投稿时间：2025-01-05 修订日期：2025-02-13 |
| DOI：10.16688/j.zwbh.2025005 |
| 中文关键词: 樱桃病毒A 全基因组 反转录环介导等温扩增 |
| 英文关键词:cherry virus A complete genome reverse transcription loop-mediated isothermal amplification (RT-LAMP) |
| 基金项目:安康市中青年科技创新领军人才计划（AK2024PD-ZQN-02）；陕西省高校青年创新团队（2023AYTD01）；陕西省教育厅协同创新中心（23JY003）；安康学院高层次人才引进专项（2023AYKCYZ02）；大学生创新训练项目（S202311397041，202411397014） |
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| 中文摘要: |
| 本研究获得了樱桃病毒A(cherry virus A, CVA)陕西安康樱桃分离物SXAK3-3和SXAK3-4的全基因组, 全长分别为7 440 nt和7 436 nt, 核苷酸一致性为80.77%。建立了CVA的反转录环介导等温扩增(reverse transcription loop-mediated isothermal amplification, RT-LAMP)检测技术并优化了反应条件和体系。该检测体系可特异检测CVA，对樱桃小果病毒2（little cherry virus 2, LChV-2）、李属坏死环斑病毒(prunus necrotic ringspot virus, PNRSV)、柑橘叶斑驳病毒（citrus leaf blotch virus, CLBV）和樱桃绿环斑驳病毒(cherry green ring mottle virus, CGRMV) 4种樱桃病毒的检测结果均为阴性，其检测灵敏度是RT-PCR的100倍。RT-LAMP检测CVA的成本低、速度快、特异性强, 在CVA抗病毒育种、苗木带毒检测、病毒早期监测等方面将发挥重要作用。 |
| 英文摘要: |
| In this study, the complete genomes of two cherry virus A (CVA) isolates from cherry in Ankang, Shaanxi province, designated SXAK3-3 and SXAK3-4, were successfully amplified. Their full lengths were 7 440 nt and 7 436 nt, respectively, sharing a nucleotide identity of 80.77%. A reverse transcription loop-mediated isothermal amplification (RT-LAMP) detection system for CVA was established and its reaction conditions and parameters were optimized. The developed RT-LAMP assay specifically detected CVA, but not four other cherry viruses: little cherry virus 2 (LChV-2), prunus necrotic ringspot virus (PNRSV), citrus leaf blotch virus (CLBV), and cherry green ring mottle virus (CGRMV). Its detection sensitivity is 100 times that of RT-PCR. Owing to its low-cost, rapidity, and strong specificity, the RT-LAMP assay provides an efficient tool for virus-resistance breeding, detection of virus-carrying seedlings, and early monitoring of CVA. |
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