Division of Biotechnology, Taiwan Agricultural Research Institute, Taichung, Taiwan.
Division of Plant Pathology, Taiwan Agricultural Research Institute, Taichung, Taiwan.
PeerJ. 2022 May 26;10:e13409. doi: 10.7717/peerj.13409. eCollection 2022.
Banana bunchy top virus (BBTV), cucumber mosaic virus (CMV) and banana streak virus (BSV) are important banana viruses, there are possible infections frequently with several viruses in field. Since the viruses are readily trasmitted in vegetative propagules, which pose a threat to banana production in banana-growing areas.
A multiplex polymerase chain reaction (PCR) protocol combined with LiquiChip analysis to identify BSV, BBTV, and CMV, with consistent amplification of plant ubiquitin (), the banana plant messenger RNA used as a procedural control. Multiplex reverse transcription (RT)-PCR amplicons were extended by allele-specific primers, followed by hybridization with carboxylated microspheres containing unique fluorescent oligonucleotides, which were detected using the LiquiChip 200 workstation.
In this study, we aimed to develop a rapid, sensitive, and simultaneous detection method for BSV, BBTV, and CMV using a bead-based multiplex assay that can be applied in routine diagnosis. We demonstrated that this detection system was extremely efficient and highly specialized for differentiating individual in a mixture of viruses while being ten times more sensitive than traditional RT-PCR. The development of this method makes it feasible to detect banana viruses in field collected leaf samples.
香蕉束顶病毒（BBTV）、黄瓜花叶病毒（CMV）和香蕉条纹病毒（BSV）是重要的香蕉病毒，田间常有几种病毒的复合侵染。由于这些病毒很容易在营养繁殖体中传播，这对香蕉种植区的香蕉生产构成了威胁。
采用多重聚合酶链反应（PCR）结合 LiquiChip 分析方法鉴定 BSV、BBTV 和 CMV，一致扩增植物泛素（），香蕉植物信使 RNA 用作程序对照。多重逆转录（RT）-PCR 扩增子用等位基因特异性引物延伸，然后与含有独特荧光寡核苷酸的羧基化微球杂交，使用 LiquiChip 200 工作站进行检测。
本研究旨在开发一种基于珠的多重检测方法，用于快速、灵敏、同时检测 BSV、BBTV 和 CMV，该方法可用于常规诊断。我们证明，该检测系统在区分混合物中的单个病毒时非常高效和高度特异，比传统 RT-PCR 灵敏 10 倍。该方法的开发使得在田间采集的叶片样本中检测香蕉病毒成为可能。
