吴凤雷,黄瑜,黄郁葱,蔡双虎,简纪常,汤菊芬

• 1:广东海洋大学水产学院广东省水产经济动物病原生物学及流行病学重点实验室暨水产经济动物病害控制广东省普通高等学校重点实验室

摘要(Abstract)：

【目的】建立一种快速、灵敏、特异的罗非鱼(Oreochromis spp)罗湖病毒定量检测方法。【方法】根据罗湖病毒节段8保守区设计一对特异性引物,建立检测罗湖病毒的SYBRGreenⅠ实时荧光定量RT-PCR方法,并对该方法的特异性、灵敏度和重复性进行了评价。【结果】标准曲线在2.5×10~8～2.5×10~0拷贝数之间有良好的线性关系(相关系数R~2=0.999),检测限为2.5×10~0个拷贝。试验内及试验间变异系数分别为0.11%～0.43%与0.54%～1.13%,重复性强;对水生动物其他病毒和细菌均无扩增反应,有很好的特异性。【结论】新建立的罗湖病毒实时荧光定量PCR检测方法特异性好、灵敏度高、重复性强,可用于TiLV的早期的快速诊断、流行病学调查和防控。

关键词(KeyWords)： 罗非鱼;罗湖病毒;检测;实时荧光定量RT-PCR;标准曲线

Abstract:

Keywords:

基金项目(Foundation): 广东省自然科学基金(2016A030313748);; 国家重点研发计划(2018YFD0900501)

作者(Author): 吴凤雷,黄瑜,黄郁葱,蔡双虎,简纪常,汤菊芬

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