To visually and rapidly detect a novel goose astrovirus (N-GoAstV) causing fatal gout in goslings, an isothermal detection method based on one-step reverse transcription loop-mediated isothermal amplification (one-step RT-LAMP) was established. The one-step RT-LAMP assay for N-GoAstV detection, using Bst 3.0 DNA polymerase with strong reverse transcription activity and primer sets targeting the opening reading frame 1b (ORF1b) of N-GoAstV, could be completed in 30 min using a water bath at 61°C; the detection results could be visually observed by adding a pH-sensitive dye containing phenol red and cresol red. The detection limit of the one-step RT-LAMP assay was 57.8 copies, which was similar to that of reverse transcription-quantitative polymerase chain reaction. The assay specifically detected N-GoAstV without any cross-reaction with other reference viruses, and this was further confirmed using enzyme digestion. These results indicated that the newly established RT-LAMP assay could accomplish reverse transcription, amplification, and visual result determination in one step, and the results obtained via this rapid and cost-effective method could be used to support disease control on farms in terms of N-GoAstV infection.

中文翻译：

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使用一步反转录环介导的等温扩增，快速直观地检测导致雏鹅致命痛风的新型星状病毒。

为了在视觉上快速检测出引起鹅雏性致命痛风的新型鹅星状病毒（N-GoAstV），建立了一种基于一步逆转录环介导的等温扩增（一步RT-LAMP）的等温检测方法。使用具有强大逆转录活性的Bst 3.0 DNA聚合酶和靶向N-GoAstV的开放阅读框1b（ORF1b）的引物对进行N-GoAstV检测的一步式RT-LAMP分析可以用水在30分钟内完成在61℃下浴；通过添加含有酚红和甲酚红的pH敏感染料，可以目视观察检测结果。一步式RT-LAMP分析的检测限为57.8拷贝，与逆转录定量聚合酶链反应的检测限相似。该测定法特异性检测了N-GoAstV，而没有与其他参考病毒发生任何交叉反应，并且使用酶消化进一步证实了这一点。这些结果表明，新建立的RT-LAMP分析可以一步完成逆转录，扩增和视觉结果的确定，通过这种快速且经济高效的方法获得的结果可用于支持农场的疾病控制。 N-GoAstV感染。