Two reverse-transcription loop-mediated isothermal amplification (RT-LAMP) assays were developed for the detection of areca palm necrotic ringspot virus (ANRSV) and areca palm necrotic spindle-spot virus (ANSSV), respectively. These two emerging viruses both induce necrotic symptoms in areca palms. The coat protein (CP) gene of ANRSV and the 9 K gene of ANSSV were used to design the respective RT-LAMP primers for the assays. Each set of four primers designed for each of these viruses was found to be highly specific in the detection of the respective targeted virus. The optimal incubation conditions for the RT-LAMP assays were 63 °C for 40 min for ANRSV and at 61 °C for 40 min for ANSSV. The sensitivity of the RT-LAMP method for each of these viruses was 10-fold greater than that of the corresponding conventional reverse-transcription polymerase chain reaction (RT-PCR). The RT-LAMP assays may be useful for the rapid early detection of ANSSV and ANRSV in commercial areca palm production.

中文翻译：

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通过逆转录环介导的等温扩增（RT-LAMP）快速检测与槟榔槟榔中的坏死症状相关的两种新兴病毒。

开发了两种逆转录环介导的等温扩增（RT-LAMP）分析方法，分别用于检测槟榔棕榈坏死环斑病毒（ANRSV）和槟榔棕榈坏死梭状斑病毒（ANSSV）。这两种新出现的病毒都在槟榔中诱发坏死症状。使用ANRSV的外壳蛋白（CP）基因和ANSSV的9 K基因来设计相应的RT-LAMP引物。发现为这些病毒中的每一种设计的每组四个引物在检测各自的靶标病毒中具有高度特异性。RT-LAMP分析的最佳孵育条件是：ANRSV为63°C 40分钟，ANSSV为61°C 40分钟。RT-LAMP方法对每种病毒的敏感性比相应的常规逆转录聚合酶链反应（RT-PCR）的敏感性高10倍。RT-LAMP分析可用于商业槟榔生产中快速早期检测ANSSV和ANRSV。