• 생명과학회지
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• 제3권4호
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• pp.209-215
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• 1993

어류의 대량폐사의 원인은 여러 가지가 있을 수 있는데 가장 중요한 것 중의 하나가 바이러스 감염에 의한 것이다. 우리나라 송어양식의 경우, 바이러스성 질병에 의한 대량폐사가 발생하여 매년 수천만마리의 치어가 폐사되어 왔는데 이 질병의 원인을 조사한 결과, 전염성 췌장 괴사 바이러스(Infectious Pencreatic Necrosis Virus ; IPNV)와 전염성 조혈기 괴사 바이러스(Infectious Hematopoietic Necrosis Virus ; IHNV)가 분리되어 이들의 대량폐사의 병원체임읠 밝혀내었다. 따라서 본 총설에서는 연어과 어류중 양식 대상종인 무지개송어 양식시 전염성과 병원성이 강해 치어기에 들어 대량폐사를 유발시키는 IHNV에 대해 최근 연구 결과를 중심으로 바이러스 분리 및 동정, 신속진단, 바이러스 백신 기초 기술 개발순으로 기술하고자 한다.

• 미생물학회지
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• 제34권1_2호
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• pp.69-73
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• 1998

한국에서 분리된 전염성 조혈괴저바이러스(infectious hematopoietic necrosis virus, IHNV)인 IHNV-PRT의 nucleocapsid(N)를 암호화하고 있는 cDNA를 클로닝하여 분석하였다. N 유전자는 391개의 아미노산으로 구성된 42.3 kDa의 분자량을 가진 단백질을 암호화하는 1,176 bp 크기의 open reading frame을 가지고 있었다. N 단백질의 아미노산 서열을 다른 외국에서 분리한 IHNV들의 N과 비교한 결과 75-90% 정도의 유사성을 보였으나 다른 종의 fish rhabdovirus인 hirame rhabdovirus(HRV) 및 viral hemorrhagic septicemia virus(VHSV)와는 각각 43%와 38%의 유사성을 보였다. 그러나 아미노산 서열 214-265의 52개의 아미노산들은 모든 종의 fish rhabdovirus간에 매우 높은 유사성을 보여 주었다.

• 한국어병학회지
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• 제31권2호
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• pp.81-85
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• 2018

Fetal bovine serum (FBS) is an essential element of cell growth and can also affect the viral replication. In this study, we tried to find out whether FBS concentration affects the viral infectivity titer of IHNV and IPNV. EPC cells were suspended with MEM supplemented with various concentrations of FBS (MEM0, MEM2, MEM5 and MEM10) and cultured in 96-well plate. Each virus was 10-fold diluted virus and inoculated in 96-well plate. The highest infectivity titer of IHNV was $10^{7.88}\;TCID_{50}/mL$ in 96-well plate using MEM5 and the lowest one was $10^{7.30}\;TCID_{50}/mL$ in 96-well plate using MEM10. The highest infectivity titer of IPNV was $10^{7.47}\;TCID_{50}/mL$ in 96-well plate using MEM5 and the lowest one was $10^{6.97}\;TCID_{50}/mL$ in 96-well plate using MEM10. This study showed that not only 0% FBS but 10% FBS leads low infectivity titer of IHNV and IPNV. Therefore, it is considered that the desirable concentration of FBS is 2% or 5% for measurement of infectivity titer of IHNV and IPNV.

• 한국어병학회지
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• 제35권2호
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• pp.157-165
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• 2022

Single-cycle viruses generated by reverse genetic technology are replication-incompetent viruses due to the elimination of gene(s) essential for viral replication, which provides a way to overcome the safety problem in attenuated viruses. Infectious hematopoietic necrosis virus (IHNV) is a major pathogen causing severe damage in cultured salmonid species. In the present study, we generated a single-cycle IHNV lacking the transmembrane and cytoplasmic domain in the G gene (rIHNV-GΔTM) and evaluated the prophylactic potential of rIHNV-GΔTM in rainbow trout (Oncorhynchus mykiss). To produce rIHNV-GΔTM, IHNV G protein-expressing Epithelioma papulosum cyprini (EPC) cells were established. However, as the efficiency of rIHNV-GΔTM production in EPC cell clones was not high, fish were immunized with a low-tittered single-cycle virus (1.5 × 10² PFU/fish). Despite the low dose, the single-cycle IHNV induced significant protection in rainbow trout against IHNV infection, suggesting high immunogenicity of rIHNV-GΔTM. No significant difference in serum ELISA titers against IHNV between the rIHNV-GΔTM immunized group and the control group suggests that the immunized dose of rIHNV-GΔTM might be too low to induce significant humoral adaptive immune responses in rainbow trout. The involvement of adaptive cellular immunity or innate immunity in the present significantly higher protection by the immunization with rIHNV-GΔTM should be further investigated to know the protection mechanism.

• 한국어병학회지
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• 제16권3호
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• pp.161-164
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• 2003

An Infectious hematopoietic necrosis virus strain (IHNV-RTK) was isolated from cultured rainbow trout at Kumi and Jechun area in Korea during 2000 and 2001. In the RT-PCR amplification with the specific primer set designed from IHNV G protein region, a 540 bp PCR product was amplified from the RTK strain. The RTK strain showed higher sequence homology with the published IHNV G protein genes (RB-76, LR-73, Col-85, and Carson-89)

• 한국수산과학회지
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• 제30권6호
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• pp.948-955
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• 1997

Immunomagnetic separation of virus coupled with .reverse transcription-polymerase chain reaction (IMS-PCR) was performed with infectious hematopoietic necrosis virus (IHNV). A DNA fragment of expected size was synthesized in the RT-PCR with total RNA extracted from IHNV inoculated CHSE-214. In a SDS-PAGE analysis, a protein band of over 70kDa was detected from non-infected cells and cells inoculated with IHNV and infectious pancreatic necrosis virus (IPNV). This protein was detected in the Western blot analysis probably because of non-specific reaction to monoclonal antibody against IHNV nucleocapsid protein. In the immunomagnetic separation, magnetic beads coated with monoclonal antibody against the IHNV nucleocapsid protein was incubated with supernatant from IHNV inoculated CHSE-214 cells. During this process, the non-specifically reacting protein could be removed by washing the magnetic bead with PBS in the presence of an external magnetic field, and viral proteins were detected from the remaining, cleaned magnetic beads. It was necessary to extract viral RNA from the captured virus particles before RT-PCR, and no DNA product was detected when the captured virus was only heated 5 min at $95^{\circ}C$. A PCR-product of expected size was synthesized from IMS-PCR with magnetic beads double coated either by goat anti-mouse IgG antibody -monoclonal antibody or streptavidin - biotin conjugated monoclonal antibody.

• 한국어병학회지
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• 제31권1호
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• pp.1-8
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• 2018

Causative agent of infectious hematopoietic necrosis (IHN) belonging to genus Novirhabdovirus (Rhabdoviridae). Economic losses caused by IHNV are serious in mainly Oncorhynchus spp. including rainbow trout O. mykiss and Atrantic salmon Salmo salar. IHNV was initially found by endemic presence in U.S. West Coast for sockeye salmon fry O. nerka and chinook salmon fry O. tshawytscha in the 1950s, and it has spread to Japan, Korea and Taiwan in the 1970s, and also to Italy and France in the 1990s. Currently, IHNV is detectable in many parts of the world, including Russia and South America. Mortality due to IHNV infection in fish with ${\leq}0.5g$ of body weight reaches 60% to 100%, while the mortality reduces by fish growing. In recent years, onset of IHNV infection has increased also in fish with large sizes. Here, we introduce molecular epidemiology and virulence changes of IHNV in East Asia, furthermore, we discuss on future prospects in IHNV researches.

• 한국어병학회지
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• 제6권2호
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• pp.87-92
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• 1993

1. 1990년 2월 강원도 삼척군에 소재한 송어양식장에서 전염성 조혈기 괴사증 증상으로 산천어 자치어가 대량폐사하였다. 2. 폐사한 산천어 치어로부터 세포배양법에 의해 전염성 조혈기 괴사증 바이러스가 분리되었다. 3. 분리된 바이러스는 구조단백의 크기와 항원성에 있어 미국에서 분리된 RB-76 바이러스주(electro-pherotype 1)와 유사하였다.

• 한국어병학회지
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• 제23권1호
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• pp.1-8
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• 2010

무지개 송어 발안란의 이동 이력과 IHNV G gene 염기서열의 계통학적 분석에 의해 IHNV의 국내 이동에 대해서 처음으로 조사되었다. IHNV RtWanju09 분리주는 IHNV RtPy91과 RtJe00의 JRt Shizuoka lineage에 속하는 것으로 나타났으며, 강원도에서 분리된 IHNV RtPy91과 RtWanju09 사이의 G gene의 유전적 차이는 1.77%였고, 약 20년간 우리나라의 JRt Shizuoka lineage에 속하는 한국 분리주들은 최대 3.03%의 유전적 차이를 나타내 계속적으로 바이러스가 진화하는 것을 알 수 있었다. 또한 전라북도 지역에서 분리된 IHNV RtWanju09 분리주는 IHNV가 오염된 강원도 지역산 발안란이 전라북도에 전파되었을 것으로 사료된다.

• Fisheries and Aquatic Sciences
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• 제4권1호
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• pp.32-38
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• 2001

Several molecular biological techniques have been used to detect virus rapidly and accurately, but these methods have limitations in the early stage of viral infection with very low concentration of virus. We compared the detection limits of IMS-PCR, Western blot and ELISA with infectious hematopoietic necrosis virus OHNV). Four antibodies, rabbit anti-IHNV polyclonal antibody, anti-IHNV nucleocapsid protein monoclonal antibody, anti-IHNV nucleocapsid protein polyclonal antibody, and anti-IHNV glycoprotein polyclonal antibody, were tested to find out the most effective antibody for each method. The detection limit with IMS- PCR was $2\times10^6$ pfu when the viral RNA was extracted before RT-PCR. In the western blot with rabbit anti­IHNV polyclonal antibody one pfu of virus could be detected. In ELISA, 10 pfu of virus particles were detected with the same antibody.