• 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.

• 생명과학회지
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• 제15권2호
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• pp.248-252
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• 2005

대하 새우양식장의 대하에 흰반점 증상을 나타내는 원인바이러스는 막대형의 이중막을 가지고 있었으며, 전자 현미경 관찰 결과 평균 크기가 $250\~300\times70\;nm$였고, 조직학적 병변은 위상피 등에서 핵이 비대해지는 것이 관찰되었다. 공격실험에서는 건강체의 새우에 많은 누적 폐사율을 보였다. 원인 바이러스 단백질은 21개의 밴드를 보였으며, 핵산 분석 결과 total 분자량은 114 kb로 나타났다.

• 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.

• 한국어업기술학회:학술대회논문집
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• 한국어업기술학회 2001년도 춘계 수산관련학회 공동학술대회발표요지집
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• pp.524-525
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• 2001

White spot syndrome virus (WSSV) causes the most serious epizootic in cultured penaeid shrimp. The epizootic started in 1992, and spread through east and south east Asia and into other shrrimp growing countries of the region. In order to circumvent the epizootic, in many Asian countries, freshwater prawn, Macrobrachium rosenbergii is being widely considered as an alternative species to marine shrimp. (omitted)

• 한국어병학회지
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• 제32권2호
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• pp.123-127
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• 2019

In this study, white spots appearing in carapace of imported white leg shrimp Litopenaeus vannamei were observed by macroscopic and microscopic examination. White spots due to white spot disease showed two types, white spots with a regular circular shape ranging from 0.4 mm to 1.7 mm and white spots with an irregular shape ranging from 0.05 mm to 2 mm. White spots due to other causes showed four types that appeared as crystalized materials in range of 100 ㎛ around the tegumental gland, as polygonal inorganic deposits ranging from 58 ㎛ to 188 ㎛, as Linear inorganic deposits ranging from 0.8 mm to 9 mm, as brownish spots ranging from 0.1 mm to 0.4 mm, and brownish spots containing melanin ranging from 0.1 mm to 2 mm. As a result, white spots occurred in imported white leg shrimp Litopenaeus vannamei were divide two type, due to white spot disease or other causes, for quarantine works.

• Journal of Microbiology and Biotechnology
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• 제11권3호
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• pp.394-398
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• 2001

Because of the great concern over the possibility of contamination from the rod-shaped nuclear virus (PRDV) from Japan and white spot virus (WSSV) from Taiwan, most eggs used in Korean shrimp farms are currently obtained from local broodstock. In addition, the screening of imported broodstock for any viral presence at the National Fisheries Research and Development Institute is also mandatory. Nonetheless, massive mortality from white spot syndrome continues in Korea. In the present study, we present an improved PCR method to use tissue-extracted DNA instead of viral DNA extracted from a purified virus based on a sucrose density gradient, and produced results within 8 h. In 1998, this modified PCR method was able to detect that diseased Penaeus japonicus were infected within 8 h. In 1998, this modified PCR method was able to detect that diseased Penaeus japonicus were infected only with PRDV, while Fenneropenaeus chinensis were infected with both PRDV and WSSV. In 1999, PRDV and WSSV were detected in F. chinensis with signs of infection, but not with WSSV alone.

• BMB Reports
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• 제37권6호
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• pp.726-734
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• 2004

White spot disease (WSD) is caused by the white spot syndrome virus (WSSV), which results in devastating losses to the shrimp farming industry around the world. However, the mechanism of virus entry and spread into the shrimp cells is unknown. A binding assay in vitro demonstrated VP28-EGFP (envelope protein VP28 fused with enhanced green fluorescence protein) binding to shrimp cells. This provides direct evidence that VP28-EGFP can bind to shrimp cells at pH 6.0 within 0.5 h. However, the protein was observed to enter the cytoplasm 3 h post-adsorption. Meanwhile, the plaque inhibition test showed that the polyclonal antibody against VP28 (a major envelope protein of WSSV) could neutralize the WSSV and block an infection with the virus. The result of competition ELISA further confirmed that the envelope protein VP28 could compete with WSSV to bind to shrimp cells. Overall, VP28 of the WSSV can bind to shrimp cells as an attachment protein, and can help the virus enter the cytoplasm.

• 한국어병학회지
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• 제10권2호
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• pp.87-95
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• 1997

새우의 갑각에 흰점을 유발하는 특징을 가진 WSBV는 Baculovirus의 일종으로 여러 종류의 새우에 높은 치사율을 보이는 병원체로서 새우양식에 막대한 피해를 주고 있다. 본 연구에서는 국내에서 양식중인 대하에 질병을 유발한 WSBV의 특성을 알아내고자 치사한 새우로부터 바이러스의 게놈을 클로닝하여 재조합클론(E3)을 분자생물학적으로 분석하였다. E3의 염기서열을 분석한 결과, 이 클론은 AcNPV를 포함한 지금까지 알려진 어떠한 바이러스와도 뚜렷한 상동성(60%)을 보이지 않아 WSBV가 기존의 바이러스와 구분되는 새로운 바이러스임을 알 수 있다. E3의 염기서열에 기초하여 한쌍의 PCR 프라이머를 작성하였다. 병든 새우로부터 분리한 DNA를 30회 증폭한 결과, 예상크기의 산물을 얻을 수 있어 이 방법은 바이러스의 감염여부를 알아낼 수 있는 진단법으로도 활용가능하다.

• 한국어병학회지
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• 제27권1호
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• pp.11-16
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• 2014

The resistance against white spot syndrome virus (WSSV) infection in wild marine crab Gaetice depressus by the immunization of a recombinant glutathione-S-transferase (GST) fused VP28 protein (GST-VP28) was evaluated. The cumulative mortalities of GST-VP28 injected groups were lower than those of the control groups at 10 days of post-challenge, and the time to death of 50% crab ($TD_{50}$) was delayed by the immunization using GST-VP28. The group boosted with GST-VP28 after 2 weeks of primary immunization clearly showed longer $TD_{50}$ than non-boosted group against challenge with WSSV. This result suggests that boosting with the antigen protein elicit stronger immune responses similar to adaptive immune responses of vertebrates. However, the short $TD_{50}$ was observed in the group challenged at 3 weeks post boosting comparing to the group challenged at 1 week post boosting. This suggests that the protective strength of immunization decreased by the time.

• 한국어병학회지
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• 제33권2호
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• pp.139-143
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• 2020

This study investigated the disinfection effect of monopersulfate (MPS) compound against white spot syndrome virus (WSSV) by bioassay using kuruma shrimp (Marsupenaeus japonicas). A WSSV stock was prepared with muscle homogenate from WSSV-infected whiteleg shrimp (Penaeus vannamei) and its lethal dose 50% endpoint (LD50) and infectious dose 50% endpoint (SID50) were respectively determined as 10-5.63 and 10-6.79 in bioassay using kuruma shrimp, followed by PCR assays. The disinfective effect of MPS compound (1.2 ppm, 2.4 ppm, 4.8 ppm) was performed by bioassay using about 10-fold higher dilution (10-4) of WSSV homogenate. The compound resulted in WSSV inactivation by a concentration-dependent manner. In addition, 4.8 ppm of MPS completely prohibited WSSV infection. To our knowledge, this study is the first report about the usefulness of MPS as a disinfectant to WSSV.