基于转录重组酶聚合酶的等温扩增与侧向流动免疫层析分析联合目视检测柑橘碎叶病毒的研究。
Development of transcription recombinase polymerase based isothermal amplification coupled with lateral flow immunochromatographic assay for visual detection of citrus tatter leaf virus.
机构信息
National Citrus Engineering and Technology Research Center, Citrus Research Institute, Southwest University, Chongqing 400712, China.
Horticultural Research Institute, Guangxi Academy of Agricultural Sciences, Nanning 530007,China.
出版信息
J Virol Methods. 2022 Nov;309:114593. doi: 10.1016/j.jviromet.2022.114593. Epub 2022 Jul 28.
The citrus tatter leaf virus (CTLV) is one of the most destructive citrus viral diseases worldwide. In this study, reverse transcription-recombinase polymerase amplification combined with a lateral flow dipstick (RT-RPA-LFD) assay for rapid visual detection of CTLV was established. The assay was performed at 35 ℃ in 27 min without specialised equipment. The RT-RPA-LFD assay showed high specificity to CTLV, and the sensitivity to CTLV was the same as that of quantitative RT-PCR at 3 × 10 copies/μL CTLV RNA transcripts. A total of 45 field tangor samples were tested using RT-RPA-LFD, RT-PCR, and RT-qPCR, and the results were consistent. The results demonstrated that the RT-RPA-LFD assay is a promising tool for rapid on-site CTLV detection.
柑橘碎叶病毒（CTLV）是世界范围内最具破坏性的柑橘病毒性疾病之一。本研究建立了一种用于快速目视检测 CTLV 的逆转录-重组酶聚合酶扩增结合侧流层析检测（RT-RPA-LFD）方法。该测定在 35℃下 27 分钟内完成，无需特殊设备。RT-RPA-LFD 测定法对 CTLV 具有高度特异性，对 CTLV 的灵敏度与定量 RT-PCR 相同，在 3×10 拷贝/μL CTLV RNA 转录本时相同。使用 RT-RPA-LFD、RT-PCR 和 RT-qPCR 对 45 个田间橘样本进行了检测，结果一致。结果表明，RT-RPA-LFD 测定法是一种快速现场 CTLV 检测的有前途的工具。
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