大口黑鲈重要病原三重PCR检测方法的建立  

Establishment of triple PCR detection method for important pathogens in largemouth bass

作　　者：林而舒 LIN Ershu(Freshwater Fisheries Research Institute of Fujian Province,Fuzhou,Fujian 350002,China)

机构地区：[1]福建省淡水水产研究所,福建福州350002

出　　处：《福建农林大学学报（自然科学版）》2024年第4期517-521,共5页Journal of Fujian Agriculture and Forestry University:Natural Science Edition

基　　金：福建省海洋服务与渔业高质量发展专项资金项目(FJHY-YYKJ-2023-2-4);福建省公益类科研院所基本科研专项(2023R1013003、2022R1014005)。

摘　　要：【目的】为同时检测患病大口黑鲈蛙病毒(Rana)、肿大细胞病毒(Mega)和弹状病毒(Rhab)的感染情况,建立了其三重PCR检测方法。【方法】设计了Rana、Mega和Rhab 3对特异性上、下游引物进行三重PCR检测,Rana、Mega和Rhab扩增片段长度分别为202、360、563 bp,并进行三重PCR反应体系的优化。【结果】建立的三重PCR方法对Rana、Mega和Rhab核酸检出限最低均可达5.3×10^(-6)μg·mL^(-1)。对临床获得的5份大口黑鲈病料进行三重PCR检测,获得2个Rana、1个Mega和1个Rhab检测阳性结果。【结论】本研究建立的大口黑鲈重要病原三重PCR检测方法,可用于养殖大口黑鲈过程中病毒病快速鉴别诊断和分子流行病学调查。【Objective】In order to simultaneously detect Ranavirus(Rana),Megalocytivirus(Mega)and Rhabdovirus(Rhab),3 important pathogens of largemouth bass(Micropterus salmoides),a triple PCR detection method was established.【Method】Three pairs of specific primers were designed for triple PCR,and the amplified fragments were 202,360 and 563 bp in length for Rana,Mega and Rhab,respectively.Reaction conditions of the triple PCR were optimized to improve the specificity and sensitivity.【Result】The detection limit of the triple PCR for Rana,Mega and Rhab nucleic acids was 5.3×10^(-6)μg·mL^(-1).Among the 5 clinical samples of largemouth bass tested by the triple PCR,2,1,and 1 samples were respectively positive for Rana,Mega and Rhab.【Conclusion】The developed triple PCR method could be used for rapid diagnosis and epidemiology investigation on virus disease of cultured largemouth bass.

关 键 词：大口黑鲈 三重PCR检测 蛙病毒 肿大细胞病毒 弹状病毒 

分 类 号：S941[农业科学—水产养殖]