naa a22 4500 510754198 CHVBK 20180411083116.0 cr unu---uuuuu 180411e20131001xx s 000 0 eng 10.1007/s13131-013-0370-9 doi (NATIONALLICENCE)springer-10.1007/s13131-013-0370-9 Diagnosis of nervous necrosis virus in orange-spotted grouper, Epinephelus coioides , by a rapid and convenient RT-PCR method [Elektronische Daten] [Yinnan Mu, Kebing Lin, Xinhua Chen, Jingqun Ao] Viral nervous necrosis (VNN) causes high mortality in marine fish, especially in the grouper, worldwide and in China. Since there is no effective vaccine or drug to deal with VNN, early detection and prevention is important to block its outbreak. In this study, a reverse transcription-polymerase chain reaction (RT-PCR) was developed for the rapid, convenient, and sensitive detection of the VNN pathogen, nervous necrosis virus (NNV), in the grouper. The whole process was completed within 3.5 h from the RNA extraction to PCR product visualization. The detection limit of this method was 200 copies of NNV RNA standard, which corresponded to 200 copies of virus particles. This RT-PCR method was specific to the NNV detection with no cross-reactivity to other fish viral disease pathogens, such as infectious pancreatic necrosis virus (IPNV), infectious hematopoietic necrosis virus (IHNV), spring viraemia of carp virus (SVCV), epizootic haematopoietic necrosis virus (EHNV), and large yellow croaker iridovirus (LYCIV). With this method, the orange-spotted grouper (Epinephelus coioides) fry from hatcheries with or without incidence of the VNN epidemic in Fujian Province were detected. The results showed that all or 93% of the fry from the two hatcheries with incidence of the epidemic were diagnosed as positive, while 40% or 25% of fry from the two hatcheries without the VNN epidemic were also detected as NNV positive, indicating that this RT-PCR method can be used for rapid, sensitive detection of NNV infection and applied in the VNN epidemic alert. The Chinese Society of Oceanography and Springer-Verlag Berlin Heidelberg, 2013 viral nervous necrosis nationallicence nervous necrosis virus nationallicence orange-spotted grouper ( Epinephelus coioides ) nationallicence reverse transcription-polymerase chain reaction nationallicence detection nationallicence Mu Yinnan Key Laboratory of Marine Biogenetic Resources, Third Institute of Oceanography, State Oceanic Administration, 361005, Xiamen, China aut Lin Kebing Fujian Fisheries Research Institute, 361000, Xiamen, China aut Chen Xinhua Key Laboratory of Marine Biogenetic Resources, Third Institute of Oceanography, State Oceanic Administration, 361005, Xiamen, China aut Ao Jingqun Key Laboratory of Marine Biogenetic Resources, Third Institute of Oceanography, State Oceanic Administration, 361005, Xiamen, China aut Acta Oceanologica Sinica Springer Berlin Heidelberg 32/10(2013-10-01), 88-92 0253-505X 32:10<88 2013 32 13131 https://doi.org/10.1007/s13131-013-0370-9 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s13131-013-0370-9 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Mu Yinnan Key Laboratory of Marine Biogenetic Resources, Third Institute of Oceanography, State Oceanic Administration, 361005, Xiamen, China aut NATIONALLICENCE 700 1- Lin Kebing Fujian Fisheries Research Institute, 361000, Xiamen, China aut NATIONALLICENCE 700 1- Chen Xinhua Key Laboratory of Marine Biogenetic Resources, Third Institute of Oceanography, State Oceanic Administration, 361005, Xiamen, China aut NATIONALLICENCE 700 1- Ao Jingqun Key Laboratory of Marine Biogenetic Resources, Third Institute of Oceanography, State Oceanic Administration, 361005, Xiamen, China aut NATIONALLICENCE 773 0- Acta Oceanologica Sinica Springer Berlin Heidelberg 32/10(2013-10-01), 88-92 0253-505X 32:10<88 2013 32 13131 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer