Dong Xuan, Chen Yujin, Lou Haoyu, Wang Guohao, Zhou Chengyan, Wang Liying, Li Xuan, Luo Jingfei, Huang Jie
State Key Laboratory of Mariculture Biobreeding and Sustainable Goods, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Laboratory for Marine Fisheries Science and Food Production Processes, Laoshan Laboratory, Key Laboratory of Maricultural Organism Disease Control, Ministry of Agriculture and Rural Affairs, Qingdao Key Laboratory of Mariculture Epidemiology and Biosecurity, Qingdao 266071, China.
Jiangsu Shufeng Aquatic Seed Industry Co., Ltd., Gaoyou 255654, China.
Animals (Basel). 2024 Feb 11;14(4):592. doi: 10.3390/ani14040592.
Infections with (EHP), infectious hypodermal and hematopoietic necrosis virus (IHHNV), and Decapod iridescent virus 1 (DIV1) pose significant challenges to the shrimp industry. Here, a melting curve-based triple real-time PCR assay based on the fluorescent dye Eva Green was established for the simultaneous detection of EHP, IHHNV, and DIV1. The assay showed high specificity, sensitivity, and reproducibility. A total of 190 clinical samples from Shandong, Jiangsu, Sichuan, Guangdong, and Hainan provinces in China were evaluated by the triple Eva Green real-time PCR assay. The positive rates of EHP, IHHNV, and DIV1 were 10.5%, 18.9%, and 44.2%, respectively. The samples were also evaluated by TaqMan qPCR assays for EHP, DIV1, and IHHNV, and the concordance rate was 100%. This illustrated that the newly developed triple Eva Green real-time PCR assay can provide an accurate method for the simultaneous detection of three shrimp pathogens.
对虾肝胰腺细小病毒（EHP）、传染性皮下和造血组织坏死病毒（IHHNV）以及十足目虹彩病毒1型（DIV1）感染给对虾养殖业带来了重大挑战。在此，基于荧光染料Eva Green建立了一种基于熔解曲线的三重实时荧光定量PCR检测方法，用于同时检测EHP、IHHNV和DIV1。该检测方法具有高特异性、高灵敏度和高重复性。采用Eva Green三重实时荧光定量PCR检测方法对来自中国山东、江苏、四川、广东和海南五省的190份临床样本进行了评估。EHP、IHHNV和DIV1的阳性率分别为10.5%、18.9%和44.2%。还采用TaqMan荧光定量PCR检测方法对样本中的EHP、DIV1和IHHNV进行了评估，符合率为100%。这表明新开发的Eva Green三重实时荧光定量PCR检测方法可为同时检测三种对虾病原体提供一种准确的方法。
