罗湖病毒RT-LAMP方法的建立与应用
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S855.3

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福建省引导性项目(2019N0030)


Establishment and Application of an RT-LAMP Method for Detecting TiLV
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    摘要:

    为快速检测罗湖病毒(tilapia lake virus,TiLV),根据TiLV编码RNA聚合酶的片段1,设计了6条特异性引物,通过对反应体系优化,建立了检测TiLV的RT-LAMP方法,并对该方法的特异性、灵敏性、重复性进行了评价。结果显示:该方法特异性好,仅能够对TiLV进行特异性扩增,对真鲷虹彩病毒等10种鱼类常见病毒扩增均呈阴性;灵敏度为2.5×10-7 ng/μL,且30 min即可得到试验结果;两组重复试验显示,该方法重复性良好,且熔解曲线相一致。结果表明,成功建立了快速检测TiLV的RT-LAMP方法。本研究为快速诊断、监测TiLV提供了技术支撑。

    Abstract:

    In order to rapidly detect tilapia lake virus(TiLV),six specific primers were designed based on the encoded segment 1 of RNA polymerase enzyme of TiLV,followed by optimization of the reaction system,an RT-LAMP method was established,then its specificity,sensitivity and reproducibility were evaluated. The results showed that the method was with good specificity and could specifically amplify TiLV only,and it was negative to 10 common fish viruses including red sea bream iridovirus;the detection limit was 2.5×10-7 ng/μL and results could be obtained in 30 minutes;and it was found that,by two repeated tests,the method was with good repeatability,and the melting curves for the two tests were consistent. Thus,an RT-LAMP method for rapid detection of TiLV was successfully established,which technically supported future rapid diagnosis and monitoring of TiLV.

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徐淑菲,林双庆,陈信忠,刘启霖,蔡绍鑫,曾韵颖,朱黄鑫,方成俊,孔凡德.罗湖病毒RT-LAMP方法的建立与应用[J].《中国动物检疫》编辑部,2021,38(12):99-103.

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  • 在线发布日期: 2021-12-03
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