Objective:To develop the rapid and efficient quantitative detection tool for nervous necrosis virus isolated from sevenband grouper Hyporhodus septemfasciatus.Methods:The viral genes of the NNV(SGYeosu08) isolated fro...Objective:To develop the rapid and efficient quantitative detection tool for nervous necrosis virus isolated from sevenband grouper Hyporhodus septemfasciatus.Methods:The viral genes of the NNV(SGYeosu08) isolated from sevenband grouper were phylogenetically analyzed.In addition,novel quantitative PCR primers based on the genomic sequence of SGYeosu08 isolate were designed and compared it with the conventional bio-assay method(TCID_(50)) using in vitro and in vivo samples.Results:The phylogenetic analysis of viral genes demonstrated the relationship of SGYeosu08 with members of red-spotted grouper nervous necrosis virus(RGNNV).The qNNV_Rl primer set(R1_F and R1_R) and the qNNV_R2 primer set(R2_F and R2_R) revealed 93%primer efficiency(regression:y=-0.2861 x + 9.9401,R^2= 0.9976)and the revealed 108%primer efficiency(regression:y=-0.3172 x + 10.0611,R^2= 0.9982),respectively.Its comparison with viral infectivity calculated by TCID_(50) method showed similar kinetic pattern at in vitro and NNV challenged fish(in vivo) samples.Conclusions:Result show that this method is rapid and efficient to diagnose NNV infection compare to traditional bioassay method(TCID_(50)).展开更多
Rock bream (Oplegnathus fasciatus) is one of the most economically valuable fish in Korea. In recent years, artificial breeding techniques with molecular and microsatellite markers have been developed to enhance rock ...Rock bream (Oplegnathus fasciatus) is one of the most economically valuable fish in Korea. In recent years, artificial breeding techniques with molecular and microsatellite markers have been developed to enhance rock bream resources. Microsatellite loci to define genetic diversity were screened in rock bream (n = 30) from Jeju areas of Korea and fifteen polymorphic microsatellite loci were newly identified and analyzed. The number of alleles per locus ranged from 9 to 34 while observed and expected heterozygosity ranged from 0.600 to 1.000 and from 0.772 to 0.977, respectively. These markers will serve as a foundation for future population genetic studies and the selective breeding technology of rock bream farming.展开更多
基金a part of the project titled'Production of diagnostic antibodies for viral diseases in aquatic animals'(Project No.20150259)funded by the Ministry of Oceans and Fisheries,Korea
文摘Objective:To develop the rapid and efficient quantitative detection tool for nervous necrosis virus isolated from sevenband grouper Hyporhodus septemfasciatus.Methods:The viral genes of the NNV(SGYeosu08) isolated from sevenband grouper were phylogenetically analyzed.In addition,novel quantitative PCR primers based on the genomic sequence of SGYeosu08 isolate were designed and compared it with the conventional bio-assay method(TCID_(50)) using in vitro and in vivo samples.Results:The phylogenetic analysis of viral genes demonstrated the relationship of SGYeosu08 with members of red-spotted grouper nervous necrosis virus(RGNNV).The qNNV_Rl primer set(R1_F and R1_R) and the qNNV_R2 primer set(R2_F and R2_R) revealed 93%primer efficiency(regression:y=-0.2861 x + 9.9401,R^2= 0.9976)and the revealed 108%primer efficiency(regression:y=-0.3172 x + 10.0611,R^2= 0.9982),respectively.Its comparison with viral infectivity calculated by TCID_(50) method showed similar kinetic pattern at in vitro and NNV challenged fish(in vivo) samples.Conclusions:Result show that this method is rapid and efficient to diagnose NNV infection compare to traditional bioassay method(TCID_(50)).
文摘Rock bream (Oplegnathus fasciatus) is one of the most economically valuable fish in Korea. In recent years, artificial breeding techniques with molecular and microsatellite markers have been developed to enhance rock bream resources. Microsatellite loci to define genetic diversity were screened in rock bream (n = 30) from Jeju areas of Korea and fifteen polymorphic microsatellite loci were newly identified and analyzed. The number of alleles per locus ranged from 9 to 34 while observed and expected heterozygosity ranged from 0.600 to 1.000 and from 0.772 to 0.977, respectively. These markers will serve as a foundation for future population genetic studies and the selective breeding technology of rock bream farming.
