Hoffmann Lucia Vieira, Branquinho Amanda Alves, Barroso Paulo Augusto Vianna, Vaslin Maite F S
Brazilian Agricultural Research Corporation (EMBRAPA), Brasília, Brazil.
Biological Science Institute, Universidade Federal de Goiás, Goiânia, Brazil.
Front Plant Sci. 2022 Jul 13;13:814119. doi: 10.3389/fpls.2022.814119. eCollection 2022.
The cotton blue disease, caused by the cotton leafroll dwarf virus (CLRDV), leads to dwarfism, leaf rolling, and production loss in susceptible cotton varieties. To develop an enzyme-linked immunosorbent assay (ELISA) test to detect the virus in cotton and weeds, peptides based on the coat protein were used to produce polyclonal (α-GQE, α-PRN, and α-INK) and monoclonal (α-GQE, α-PRN, and α-NKF) antibodies. All six were tested as capture antibodies, and polyclonal α-GQE and the monocle onal α-NKF were labeled with the enzyme alkaline phosphatase and used as detection antibodies for a double antibody sandwich (DAS) ELISA method, in which p-nitrophenyl phosphate was added and measured by absorbance at 405 nm. The DAS-ELISA sandwich was efficient in discriminating between healthy and diseased plant extracts. The ELISA methodology detected the virus in the weeds sp., which was confirmed by RT-PCR. The monoclonal antibodies may be used to develop other diagnostic procedures.
由棉花卷叶矮化病毒（CLRDV）引起的棉花蓝病，会导致易感棉花品种出现矮化、卷叶和产量损失。为了开发一种酶联免疫吸附测定（ELISA）试验来检测棉花和杂草中的该病毒，基于外壳蛋白的肽被用于制备多克隆抗体（α-GQE、α-PRN和α-INK）和单克隆抗体（α-GQE、α-PRN和α-NKF）。所有六种抗体都作为捕获抗体进行了测试，多克隆α-GQE和单克隆α-NKF用碱性磷酸酶进行标记，并用作双抗体夹心（DAS）ELISA方法的检测抗体，其中加入对硝基苯磷酸并通过在405nm处的吸光度进行测量。DAS-ELISA夹心在区分健康和患病植物提取物方面很有效。ELISA方法在杂草中检测到了该病毒，这通过逆转录聚合酶链反应（RT-PCR）得到了证实。单克隆抗体可用于开发其他诊断程序。
