• 한국어병학회지
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• 제24권2호
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• pp.47-51
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• 2011

The present study demonstrated lymphocystis disease virus (LCDV) propagation through cytopathic effects (CPE) formation and LCDV detection in olive flounder fin (FFN) cells by polymerase chain reaction (PCR) and fluorescent antibody technique (FAT) methods. Tissue filtrates from the cluster cells produced CPE in FFN cells, which initially cells became enlarged and gradually underwent fusion en masse. Infectivity of culture grown LCDV using the FFN cells reached $10^{2.3}$ $TCID_{50}$/ml at 4 days post infection and the highest titer was measured $10^{6.5}$ $TCID_{50}$/ml at 12 days. The viral DNA was detected in the cell culture supernatants showing CPE and the CPE cells by PCR. Antigen specific strong fluorescence reacting with monoclonal antibody against the virus revealed the presence of viral antigen in the cytoplasm of infected FFN cells. These results suggest that the FFN cell line originated from the olive flounder has a susceptibility of the LCDV.

• 한국어병학회지
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• 제31권1호
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• pp.49-55
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• 2018

Tendency of viral hemorrhagic septicemia virus (VHSV) detection from 2001 to 2016 in Korea was studied based on 15 reported cases. Since the VHSV was first detected from cultured olive flounder (Paralichthys olivaceus) in Pohang in 2001, it has been continuously reported from olive flounder farms in various regions of the Korean coastal area. So far, the virus has been detected from 2 farmed fishes, 12 wild marine fishes and 2 marine bivalves. All the 67 isolates were belong to VHSV genotype IVa. The predisposing factor analysis from different olive flounder farms revealed that the VHSV were highly detected from the juveniles under 40 g in body weight, in the temperature range from 9.5 to $17^{\circ}C$ and during the period of March to June. Therefore, we recommend that farmers, need to exercise caution against VHSV infection in Spring.

• 한국어병학회지
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• 제32권1호
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• pp.45-48
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• 2019

We developed and subsequently characterized mouse monoclonal antibodies (MAbs) against recombinant VP28 structural protein (rVP28) of white spot syndrome virus (WSSV). We established six hybridoma clones secreting MAbs against rVP28: 15A11, 20G6, 31H2, 34H6, 38D1 and 43A1. All six MAbs recognized the 25 kDa of protein in gill homogenates of WSSV-infected shrimp by western blot analysis, while no reactivity was observed in gill homogenates of normal shrimp. Moreover, high enzyme-linked immunosorbent assay (ELISA) optical density (OD) values (0.8-2.68) were observed in the hemolymphs from WSSV-infected shrimp, while low OD values (less than 0.24) were recorded in the hemolymphs from normal shrimp, by using these six MAbs produced in this study. These results suggest that these six MAbs are useful for the detection of WSSV.

• 한국어병학회지
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• 제35권2호
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• pp.241-246
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• 2022

White spot syndrome virus (WSSV) is a prevalent and virulent pathogen affecting cultured whiteleg shrimp (Litopenaeus vannamei) in Korea. In this study, seven monoclonal antibodies (mAbs) (10A12, 16C3, 17G4, 21G5, 22C4, 23B6 and 24G6) were produced by using purified WSSV. The reactivity of these mAbs was analysed by Western blot (WB), indirect immunofluorescence (IIF), and lateral flow immunochromatographic assay (LFIA). WB analysis demonstrated that three mAbs (17G4, 22C4, and 23B6) reacted specifically to VP28 with an approximate molecular weight of 24 kDa, mAb 16C3 reacted with approximately 17 kDa. IIF analysis demonstrated specific fluorescence signals on gill tissues of WSSV-infected shrimp, with five mAbs (10A12, 16C3, 22C4, 23B6, and 24G6), pleopods from WSSV-infected shrimp were used for LFIA, where, two mAbs (21G5 and 22C4) exhibited positive reaction. In conclusion, it can be inferred that the mAbs usage and specificity depends on the nature of assay used for diagnosis.

• 한국어병학회지
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• 제20권3호
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• pp.201-209
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• 2007

Viral Hemorrhagic Septicemia Virus (VHSV)는 유럽과 북미지역의 담수 무지개송어에 심각한 바이러스성 질병을 야기하는 원인체였으나 담수어류 뿐만 아니라 해산어류에서도 심각한 질병을 유발하며 최근 일본과 우리나라 양식 넙치에서 VHSV에 의한 대량 폐사 발생이 보고되고 있다. 본 연구는 VHS에 대한 예방학적 접근의 일환으로서 자연산 어류로부터의 VHSV 검출 및 지리적 분포, 보균 어종에 대한 유전학적인 조사를 목적으로 2003년과, 2005년도에 동중국해역 3지점, 서해안 5지점 그리고 남해안 5지점에서 어류를 채집하였다. RT-PCR을 이용한 연구결과 자연산 해산어류 160마리 중 12종 17마리에서 VHSV가 검출되어 검출율 10.6%를 보였으며 조사된 12종의 어류 중 특히 상어목에서 가장 높은 검출율을 보였다. VHSV G gene을 이용한 계통분류학적 분석결과 염기서열 분석에서 양식산어류의 분리주 (KVHS01-1)와 98.4%～99%, 아미노산 분석에서 97～99% 유사성을 나타내어 본 연구에서 조사되어진 자연산 어류에서 분리된 VHSV는 GenogroupⅠ에 속하였다.

• 한국어병학회지
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• 제32권2호
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• pp.59-67
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• 2019

본 연구에서는 넙치(Paralichthys olivaceus)로부터 분리한 바이러스성출혈성패혈증바이러스(viral hemorrhagic septicemia virus, VHSV, genotype IVa)에 대한 단클론 항체(monoclonal antibody, MAb)를 개발하였다. VHSV에 대한 항체를 생산하는 총 5개의 hybridoma clone을 생산하였다. 4개의 MAbs (2C10, 18H4, 23H6, 30B7)는 glycoprotein을 인식하였고, MAb 15E10은 nucleocapsid protein을 인식하였다. 5개의 MAbs는 western blot 상에서 VHSV에 감염된 세포와 넙치시료에 반응하였으나, 정상 세포와 넙치시료에는 반응하지 않았다. 또한 ELISA상에서 VHSV에만 반응하였고 6종의 어류바이러스(IHNV, HIRRV, SVCV, IPNV, MABV, NNV)에는 반응하지 않았다. 이상의 결과, 본 연구에서 제작된 MAbs는 VHSV에만 특이적으로 반응하는 것이 확인되어 VHSV를 검사하는데 사용될 수 있을 것으로 사료된다.

• 한국어병학회지
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• 제8권2호
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• pp.91-98
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• 1995

최근 남해안 일대의 육상 양식장에서 사육중이던 넙치(Paralichthy olivaceus) 치어가 폐사하여 조사한 결과 3개 양식장에서 바이러스가 분리되었다. 분리된 바이러스들은 모두 외막이 없는 정육면체 모양이었으며 50~55mm 정도의 크기를 지녔다. 전기영동상에서 RNA와 구조 단백질의 patterns를 확인하고, IPNV에 대한 항혈청을 사용하여 중화실험을 수행한 결과, 분리한 세 바이러스는 birnavirus인 IPNV와 매우 유사함이 밝혀졌다. 특히 분리 바이러스중 CS는 IPNV의 AB 혈청형과 DS와 YJ는 SP 혈청형과 유사하였다.

• 한국수산과학회지
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• 제50권5호
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• pp.547-552
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• 2017

The viral hemorrhagic septicemia virus (VHSV) has an extensive host range, and infects farmed and wild fish inhabiting both freshwater and marine ecosystems. Enzyme-linked immunosorbent assay (ELISA) is highly useful in diagnosing viral hemorrhagic septicemia. However, ELISA shows high, non-specific background reaction with fish antibodies. In this study, we optimized the antigen and antibody concentrations used for detecting specific antibodies in VHSV-infected olive flounder to reduce non-specific binding, and improve the sensitivity of ELISA. The results suggested that OD (optical Density) values were valid when ELISA was performed with $0.1{\mu}g/well$ of virus, involving blocking with blocking buffer (Roth, Roti-Block), 1:300-1:600 dilution with flounder antisera, and 1:1000 dilution with anti-flounder IgM and HRP-conjugated goat anti-mouse IgG for detecting the VHSV antibody in flounder sera. Furthermore, 11 different VHSV strains isolated in Korea from 2012 to 2016 were used to infect the fish. The results showed no correlation between viral pathogenicity and antibody production. This research is a basic study on the application of antibody detection in the diagnosis of viral hemorrhagic septicemia in the olive flounder.

• 한국어병학회지
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• 제22권3호
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• pp.229-237
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• 2009

The causative agent for the tunic softness syndrome of the cultured ascidian Halocynthia roretzi from Jan 1999 to Feb 2009 was identified using virus isolation and polymerase chain reaction (PCR). The pathogenicity of the isolated virus MABV UR-1 strain was determined by experimental infection trials. The cytopathic effects was observed in CHSE-214 cell line at a level 5.1% (4/78) in normal ascidian and 1.8% in abnormal ascidian showing tunic softness syndrome signs. MABV gene was detected in 16.8% (18/107) of normal and 13.1% (5/38) of abnormal organisms by PCR. The ratio of MABV isolation and gene detection was similar level in normal and soft tunic diseased ascidian. Based on the VP2/NS junction region sequences, eight strains of virus isolated from ascidian, were included in the same genogroup with MABV which is originally isolated in wide ranges of marine fish and shellfish species. The UR-1 strain caused 60% mortality (36.5% mortality in control group) by immersion infection and 37% mortality (same mortality in control group) in injection infection indicating no significant differences in infected and control groups. These results suggest that ascidian can act as reservoir of the MABV, and this virus is not directly related with the ascidian mortality.

• 한국양식학회:학술대회논문집
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• 한국양식학회 2001년도 추계 한국양식학회 학술대회
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• pp.227.2-228
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• 2001