SUCCESS STORY

Development of sensitive and specific diagnostic tools for nodavirus

infection in fish

RT-PCR assay for the diagnosis of Viral Nervous Necrosis (VNN) in fish

A RT-PCR based diagnostic technique was developed to detect the carrier

status of viral nervous necrosis virus caused by betanodavirus, a RNA virus in fish.
Viral Nervous Necrosis (VNN) also known as Viral Encephalopathy and
Retinopathy (VER), a highly pathogenic viral disease affecting a variety of cultured
marine fishes has world wide occurrence. The mortality reach as high as 80-100%
in larval / juvenile stages of fish, but the broodstock fishes often act as sub-clinical
carriers. The etiological agent, betanodaviruses are small (25-34 nm) non-
enveloped, icosahedral viruses with a bipartite, positive sense RNA genome. The
virus localizes in the brain, spinal cord, and retina of the affected fish, which
exhibit erratic swimming behavior and a range of neurological abnormalities,
including vacuolization and cellular necrosis in the central nervous system and
retina.
Diagnosis is often difficult due to the sub clinical infection and hence a need
was felt to develop a nucleic acid based molecular technique for the diagnosis of
VNN in broodstock fish and the clinically affected fish. A RT-PCR based diagnostic
tool was developed based on the partial nucleotide sequence of betanodavirus
isolates from India and has been tested in many fish species including seabass,
mullet, grouper and fresh water ornamental fishes.
The developed protocol allows the user to rapidly interpret the disease
status of the fish either by a single step or by a nested RT-PCR. The test results
are comparable to similar kits developed elsewhere. The indigenous prototype kit
has been optimized, validated under laboratory conditions using samples drawn
from different parts of the country. The technology was perfected using different
reference strains and is ready for commercialization. The developed prototype kit
would help in the detection of clinical as well as sub clinical detection of virus in
many fish species used for culture.
This diagnostic method has been developed under a collaborative research
project entitled, “Characterization and development of diagnostics for viral
nervous necrosis in seabass (Lates calcarifer) and mullet (Mugil
cephalus)” at Central Institute of Brackishwater Aquaculture (Chennai) and
C.Abdul Hakeem College (Melvisharam). The project was funded by Department
of Biotechnology, Ministry of Science and Technology, Govt. of India (New Delhi).
Dr.K.P.Jithendran
Principal Investigator