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Genome based quantification of VHSV in multiple organs of infected olive flounder (Paralichthys olivaceus) using real-time PCR.
Genes & Genomics ( IF 1.6 ) Pub Date : 2020-05-24 , DOI: 10.1007/s13258-020-00951-7
Hyunsu Kim 1 , Kyung-Wan Baek 1, 2 , Eun-Ji Ko 1 , Nguyen Thanh Luan 3 , Yunjin Lim 3 , Heyong Jin Roh 3 , Nameun Kim 3 , Do-Hyung Kim 3 , Ahran Kim 4 , Yung Hyun Choi 5 , Suhkmann Kim 6 , Heui-Soo Kim 7 , Mee Sun Ock 1 , Hee-Jae Cha 1
Affiliation  

BACKGROUND Viral hemorrhagic septicemia (VHS) is a serious viral disease that infects the olive flounder in South Korea. The Korean aquaculture industry experienced an economic loss caused by the high infectivity and mortality. OBJECTIVE This study aimed to evaluate the infection density of VHSV in various organs of the olive flounder including spleen, liver, kidney, stomach, esophagus, intestine, gill, muscle, heart, and brain. Olive flounders were collected from a local fish farm and injected subcutaneously with 106 PFU/fish. METHODS Each 15 fish were sampled at 0, 3, and 7 days post challenge (dpc), respectively, to perform quantitative analysis of VHSV using SYBR-green based real-time PCR in various tissues including spleen, liver, head-kidney, body-kidney, muscle, esophagus, stomach, intestine, gill, and brain. RESULTS Organs infected with VHSV were obtained after 3 and 7 days. Each organs were examined for viral infection using real-time PCR. The data obtained from this experiment revealed copy numbers higher than 10 copies per 100 ng cDNA in the spleen (15.26 ± 3.11 copies/100 ng of cDNA), muscle (11.24 ± 2.25 copies), and gill (14.23 ± 6.26 copies), but lower in liver, head-kidney, body-kidney, esophagus, brain and stomach. CONCLUSION The present study, together with previous data, demonstrated that the gill, spleen, and muscle are the major target organs of VHSV in olive flounder. Therefore, central monitoring of spleen, gill and muscle should be considered and might be necessary if anti-VHSV treatment is to be successful in infected olive flounder.

中文翻译:

使用实时PCR基于基因组的VHSV在感染的橄榄比目鱼(Paralichthys olivaceus)多个器官中的定量。

背景技术病毒性出血性败血病(VHS)是一种严重的病毒性疾病,感染了韩国的橄榄比目鱼。由于高传染性和高死亡率,韩国水产养殖业遭受了经济损失。目的本研究旨在评估VHSV在比目鱼的各个器官中的感染密度,包括脾脏,肝脏,肾脏,胃,食道,肠、,、肌肉,心脏和大脑。从当地的一个养鱼场收集了比目鱼,皮下注射了106 PFU /鱼。方法分别在攻击后0天,3天和7天对每条15条鱼进行采样(dpc),以基于SYBR-green的实时PCR对包括脾脏,肝脏,头肾,身体在内的各种组织进行VHSV定量分析-肾脏,肌肉,食道,胃,肠,g和大脑。结果3、7天后获得感染VHSV的器官。使用实时PCR检查每个器官的病毒感染。从该实验获得的数据显示,脾脏(15.26±3.11拷贝/ 100 ng cDNA),肌肉(11.24±2.25拷贝)和腮(14.23±6.26拷贝)中每100 ng cDNA拷贝数高于10拷贝。肝脏,头部肾脏,身体肾脏,食道,脑部和胃部较低。结论本研究与以前的数据一起证明,ill,脾和肌肉是橄榄比目鱼VHSV的主要靶器官。因此,应考虑对脾脏,腮和肌肉进行集中监控,如果要在感染的比目鱼中成功进行抗VHSV治疗,可能有必要进行集中监控。使用实时PCR检查每个器官的病毒感染。从该实验获得的数据表明,脾脏(15.26±3.11拷贝/ 100 ng cDNA),肌肉(11.24±2.25拷贝)和腮(14.23±6.26拷贝)中每100 ng cDNA的拷贝数高于10拷贝。肝脏,头部肾脏,身体肾脏,食道,脑部和胃部较低。结论本研究与以前的数据一起证明,ill,脾和肌肉是橄榄比目鱼VHSV的主要靶器官。因此,应考虑对脾脏,腮和肌肉进行集中监控,如果要在感染的比目鱼中成功进行抗VHSV治疗,可能有必要进行集中监控。使用实时PCR检查每个器官的病毒感染。从该实验获得的数据显示,脾脏(15.26±3.11拷贝/ 100 ng cDNA),肌肉(11.24±2.25拷贝)和腮(14.23±6.26拷贝)中每100 ng cDNA拷贝数高于10拷贝。肝脏,头部肾脏,身体肾脏,食道,脑部和胃部较低。结论本研究与以前的数据一起证明,ill,脾和肌肉是橄榄比目鱼VHSV的主要靶器官。因此,应考虑对脾脏,腮和肌肉进行集中监控,如果要在感染的比目鱼中成功进行抗VHSV治疗,可能有必要进行集中监控。24±2.25份)和g(14.23±6.26份),但在肝脏,头部肾脏,身体肾脏,食道,脑和胃中较低。结论本研究与以前的数据一起证明,ill,脾和肌肉是橄榄比目鱼VHSV的主要靶器官。因此,应考虑对脾脏,腮和肌肉进行集中监控,如果要在感染的比目鱼中成功进行抗VHSV治疗,可能有必要进行集中监控。24±2.25份)和g(14.23±6.26份),但在肝脏,头部肾脏,身体肾脏,食道,脑和胃中较低。结论本研究与以前的数据一起证明,ill,脾和肌肉是橄榄比目鱼VHSV的主要靶器官。因此,应考虑对脾脏,腮和肌肉进行集中监控,如果要在感染的比目鱼中成功进行抗VHSV治疗,可能有必要进行集中监控。
更新日期:2020-05-24
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