Abstract Abaca bunchy top virus (ABTV) and banana bunchy top virus (BBTV) are pathogens that infect abaca in the Philippines causing significant reduction in abaca fiber yield and quality. In this study, a sensitive loop-mediated isothermal amplification (LAMP) assay was developed for detecting ABTV and BBTV in abaca, Musa textilis Nee. Five primer sets were designed and tested to detect ABTV while three primer sets were designed and tested to detect BBTV. Out of which only one primer set for each virus was specific to their respective targets that is DNA-N for ABTV and DNA-S for BBTV for successful LAMP based detection applications. The concentrations of Bst DNA polymerase, dNTPs, incubation time and temperature were optimized for both viruses using either GelRed™ or SYBR™ Green I as detection dyes. The ABTV and BBTV LAMP assays were more sensitive than conventional PCR and were able to detect ABTV or BBTV in asymptomatic abaca samples. The developed LAMP protocols provide a simple, fast and accurate detection method that will be indispensable for the production of ABTV- and BBTV-free abaca tissue cultured plantlets. This is the first report of a LAMP-based assay for detection of ABTV and BBTV in abaca.

中文翻译：

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环介导等温扩增 (LAMP) 检测用于检测蕉麻中的蕉麻束顶病毒和香蕉束顶病毒

摘要 蕉麻束顶病毒 (ABTV) 和香蕉束顶病毒 (BBTV) 是感染菲律宾蕉麻的病原体，导致蕉麻纤维产量和质量显着下降。在这项研究中，开发了一种灵敏的环介导等温扩增 (LAMP) 测定法，用于检测蕉麻、Musa textilis Nee 中的 ABTV 和 BBTV。设计并测试了五个引物组来检测 ABTV，而设计和测试三个引物组来检测 BBTV。其中每种病毒只有一个引物组针对它们各自的目标，即 ABTV 的 DNA-N 和 BBTV 的 DNA-S，用于成功的基于 LAMP 的检测应用。使用 GelRed™ 或 SYBR™ Green I 作为检测染料，针对两种病毒优化了 Bst DNA 聚合酶的浓度、dNTP、孵育时间和温度。ABTV 和 BBTV LAMP 检测比常规 PCR 更敏感，并且能够检测无症状蕉麻样品中的 ABTV 或 BBTV。开发的 LAMP 协议提供了一种简单、快速和准确的检测方法，这对于生产无 ABTV 和 BBTV 的蕉麻组织培养苗是必不可少的。这是用于检测蕉麻中 ABTV 和 BBTV 的基于 LAMP 的测定的第一份报告。