用多重聚合酶链反应鉴定传染性脾肾坏死病毒和病毒性神经坏死在海鱼中的感染

Hidayati Kumalasari, P. E. Sudaryatma, Artanti Tri Lestari, Wahyu Nurlita, W. Nugraha, Nur Hasanah, Ida Ayu Mirah Meliana Dewi, Ni Putu Arya Shintya Anggraeni
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摘要

印度尼西亚是一个海洋国家,拥有广阔的海洋面积和海岸线,海洋物种多样性达到世界鱼类的37%。这使得印度尼西亚在适宜气候的支持下具有水产养殖潜力。然而,由传染性脾肾坏死病毒(ISKNV)和病毒性神经坏死病毒(VNN)引起的病毒性疾病的挑战将阻碍海洋水产养殖,并造成巨大的经济损失。因此,有必要开展高效、快速、精确、准确的早期检测方法来识别这些病毒性疾病。本文建立了一种多重pcr (mPCR)方法,可以同时检测ISKNV和VNN。结果表明,该方法采用MCP和CP基因靶引物分别检测ISKNV和VNN,具有较高的灵敏度和特异性。经mPCR检测的353份海鱼中,ISKNV和VNN单次感染阳性分别为14份和2份。我们还发现18条石斑鱼同时感染了这些病毒。结果表明,该方法具有效率高、速度快、精度高等特点。这一发现可以应用于ISKNV和VNN领域的实验室测试或早期检测系统。
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Infection of Infectious Spleen and Kidney Necrosis Virus and Viral Nervouse Necrosis in Seawater Fish Identifed Using Multiplex Polymerase Chain Reaction
Indonesia become a maritime country with large of ocean area and beach line with marine species diversity reaches 37% of the world’s fish. This makes Indonesia have aquaculture potential that supported by an appropriate climate. However, the challenge of viral diseases caused by Infectious Spleen and Kidney Necrosis Virus (ISKNV) and Viral Nervous Necrosis (VNN) would hamper marine aquaculture and causing huge economic losses. Therefore, it is necessary to carry out early detection methods that are efficient, fast, precise and accurate to identifying these viral disease. Here we developed a multiplex-PCR (mPCR) a method that can detect simultaneously of the ISKNV and VNN. The results of this method showed high sensitivity and specificity by using MCP and CP gene target primers to detect ISKNV and VNN, respectively. From a total of 353 samples of seawater fish examined by mPCR, positive results following of single infection of ISKNV and VNN were 14 and 2, respectively. We also found that 18 grouper fish was co-infection with these viruses. From these results, it shows that the mPCR method developed has efficiency, faster and high accuracy. This founding was possible to be applied in laboratory testing or early detection system in the field for ISKNV and VNN. 
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