چکیده (انگلیسی):

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 (TCID50) 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_R1 primer set (R1_F and R1_R) and the qNNV_R2 primer set (R2_F and R2_R)
revealed 93% primer efficiency (regression: y = −0.2861x + 9.9401, R2 = 0.9976) and the
revealed 108% primer efficiency (regression: y = −0.3172x + 10.0611, R2 = 0.9982),
respectively. Its comparison with viral infectivity calculated by TCID50 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 (TCID50).