Viral hemorrhagic septicemia virus（VHSV） and marine birnavirus（MABV） are the causative pathogens for some of the most explosive epidemics of emerging viral diseases in many Asian countries, leading to huge economi...Viral hemorrhagic septicemia virus（VHSV） and marine birnavirus（MABV） are the causative pathogens for some of the most explosive epidemics of emerging viral diseases in many Asian countries, leading to huge economic losses in aquaculture. Rapid molecular detection for surveillance or diagnosis has been a critical component in reducing the prevalence of pathogen infection. The loop-mediated isothermal amplification（LAMP） of DNA is currently one of the most commonly used molecular diagnostic tools, as it is simple, quick, and easy to amplify target DNA under isothermal conditions. In the present study, a novel and highly specific LAMP assay for the sensitive and rapid detection of VHSV and MABV infection in fish was developed. Using a set of synthesized primers matching a specific region of the genome, the efficiency and specificity of the LAMP assay were optimized in terms of the reaction temperature and DNA polymerase concentration, as they are the main determinants of the sensitivity and specificity of the LAMP assay. In particular, we demonstrated that our assay could be applied to efficient detection of VHSV and MABV infection in the wild fish, Paralichthys olivaceus. Our results demonstrate the simplicity and convenience of this method for the detection of viral infection in aquatic organisms.展开更多
In order to elucidate the state of diseases, especially viral diseases, and to prevent viral diseases from occurring in olive flounder hatcheries, a range of studies, including epidemiological study, were performed fr...In order to elucidate the state of diseases, especially viral diseases, and to prevent viral diseases from occurring in olive flounder hatcheries, a range of studies, including epidemiological study, were performed from 1997 to 2003. The location of the hatcheries investigated includes several representative sites in the east （Kangnung, Uljin, Pohang, Yangsan, Ulsan, Pusan）, south （Wando, Changheung, Goheung, Yeosu, Namhae, Tongyeong, Geoje, Jeju） and west （Seosan, Kunsan, Gochang, Yeongkwang, Mokpo, Chindo） costal areas of the Korea Peninsula. A total of 2000 cases have been examined in 7 years, in which mortality caused by viral agents accounts for 22%, or 446 cases. Mortalities associated with viral infection considerably increased from 14% in 1997 to 27% in 2003. A variety of viral diseases were observed, and the occurrences of viral epidermal hyperplasia, viral ascites and viral deformity, viral nervous necrosis, and hirame rhabdoviral disease are 14%, 51%, 25%, and 8% respectively. By investigating the viral infection of broodstock flounder, the infection rate of marine birnavirus （MABV） in hatcheries was identified to be approximately 30%, therefore, it is highly necessary to acquire and keep non-infected broodstock fishes.展开更多
基金The grants from the Korea Institute of Ocean Science and Technology under contract No.PE99315
文摘Viral hemorrhagic septicemia virus（VHSV） and marine birnavirus（MABV） are the causative pathogens for some of the most explosive epidemics of emerging viral diseases in many Asian countries, leading to huge economic losses in aquaculture. Rapid molecular detection for surveillance or diagnosis has been a critical component in reducing the prevalence of pathogen infection. The loop-mediated isothermal amplification（LAMP） of DNA is currently one of the most commonly used molecular diagnostic tools, as it is simple, quick, and easy to amplify target DNA under isothermal conditions. In the present study, a novel and highly specific LAMP assay for the sensitive and rapid detection of VHSV and MABV infection in fish was developed. Using a set of synthesized primers matching a specific region of the genome, the efficiency and specificity of the LAMP assay were optimized in terms of the reaction temperature and DNA polymerase concentration, as they are the main determinants of the sensitivity and specificity of the LAMP assay. In particular, we demonstrated that our assay could be applied to efficient detection of VHSV and MABV infection in the wild fish, Paralichthys olivaceus. Our results demonstrate the simplicity and convenience of this method for the detection of viral infection in aquatic organisms.
文摘In order to elucidate the state of diseases, especially viral diseases, and to prevent viral diseases from occurring in olive flounder hatcheries, a range of studies, including epidemiological study, were performed from 1997 to 2003. The location of the hatcheries investigated includes several representative sites in the east （Kangnung, Uljin, Pohang, Yangsan, Ulsan, Pusan）, south （Wando, Changheung, Goheung, Yeosu, Namhae, Tongyeong, Geoje, Jeju） and west （Seosan, Kunsan, Gochang, Yeongkwang, Mokpo, Chindo） costal areas of the Korea Peninsula. A total of 2000 cases have been examined in 7 years, in which mortality caused by viral agents accounts for 22%, or 446 cases. Mortalities associated with viral infection considerably increased from 14% in 1997 to 27% in 2003. A variety of viral diseases were observed, and the occurrences of viral epidermal hyperplasia, viral ascites and viral deformity, viral nervous necrosis, and hirame rhabdoviral disease are 14%, 51%, 25%, and 8% respectively. By investigating the viral infection of broodstock flounder, the infection rate of marine birnavirus （MABV） in hatcheries was identified to be approximately 30%, therefore, it is highly necessary to acquire and keep non-infected broodstock fishes.
