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Infectious hematopoietic necrosis virus (IHNV)에 대한 단클론 항체 생산
공경희 ( Kyoung-hui Kong ),오명주 ( Myung-joo Oh ),김춘섭 ( Choon-sup Kim ),김위식 ( Wi-sik Kim ) 한국어병학회 2023 한국어병학회지 Vol.36 No.2
Infectious hematopoietic necrosis virus (IHNV) is s significant viral pathogen affecting cultured rainbow trout (Oncorhynchus mykiss) in Korea. In this study, five monoclonal antibodies (mAbs) (IHNV-1, 2, 3, 4, and 5) were produced using purified IHNV. Reactivities of these mAbs were analyzed by western blot (WB), enzyme-linked immunosorbent assay (ELISA), and indirect fluorescent antibody test (IFAT). These mAbs recognized glycoprotein (69 kDa, IHNV-1), nucleocapsid protein (39 kDa, IHNV-3, 4, and 5), or phosphoprotein (27 kDa, IHNV-2) of IHNV by WB analysis. ELISA results indicated that these five mAbs were specific to IHNV without showing any cross-reactivity against other fish viruses (hirame rhabdovirus, infectious pancreatic necrosis virus, and viral hemorrhagic septicemia virus). IFAT demonstrated specific fluorescence signals of IHNV-infected epithelioma papulosum cyprini (EPC) cells, whereas no reactivity of normal EPC cells was observed. These mAbs can be very useful for immuno-diagnosis of IHNV infection.
Marine birnavirus (MABV)에 대한 단클론 항체 생산
공경희 ( Kyoung-hui Kong ),오명주 ( Myung-joo Oh ),김위식 ( Wi-sik Kim ) 한국어병학회 2020 한국어병학회지 Vol.33 No.2
We developed and subsequently characterized mouse monoclonal antibodies (mAbs) against marine birnavirus (MABV). Eight hybridoma clones secreting mAbs against MABV were established. All eight mAbs (8G6, 11C3, 15E3, 17H6, 32A6, 35A7, 38B5, and 47E3) were reacted with viral protein 3 of MABV in MABV-infected CHSE-214, whereas, no reactivity was observed in normal CHSE-214 by western blot analysis. Moreover, these eight mAbs were strongly reacted with MABV, and no cross-reactivity has been observed against other five fish viruses (hirame rhabdovirus, infectious hematopoietic necrosis virus, nervous necrosis virus, spring viraemia of carp virus, and viral hemorrhagic septicemia virus), although five mAb (11C3, 15E3, 17H6, 32A6, and 38B5) reacted with both MABV and infectious pancreatic necrosis virus by enzyme linked immunosorbent assay (ELISA). These results indicate that the mAbs can be of value in MABV detection.
VHS (viral hemorrhagic septicemia)의 원인병원체인 VHSV (genotype IVa)에 대한 단클론 항체 개발
공경희 ( Kyoung-hui Kong ),오명주 ( Myung-joo Oh ),장민석 ( Min-seok Jang ),김춘섭 ( Choon-sup Kim ),김위식 ( Wi-sik Kim ) 한국어병학회 2019 한국어병학회지 Vol.32 No.2
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 있을것으로 사료된다. We developed and subsequently characterized mouse antibodies (MAbs) against viral hemorrhagic septicemia virus (VHSV, genotype IVa), the causative agent of VHS. Five hybridoma clones secreting MAbs against VHSV were established. The MAbs recognized the glycoprotein (MAbs 2C10, 18H4, 23H6, and 30B7) and nucleocapsid protein (15E10) of VHSV by western blot analysis. All five MAbs reacted with VHSV-infected cells and tissue homogenates of VHSV-infected olive flounder (Paralichthys olivaceus) by western blot analysis. Whereas, no reactivity was observed in normal cells and tissue homogenates of normal olive flounder. Moreover, these MAbs reacted with VHSV, but did not react with other fish viruses (infectious hematopoietic necrosis virus, hirame rhabdovirus, spring viraemia of carp virus, infectious pancreatic necrosis virus, marine birnavirus, and nervous necrosis virus) by enzyme linked immunosorbent assay (ELISA). These results indicate that the MAbs are specific to VHSV and can be of value in VHSV detection.
흰다리새우(Litopenaeus vannamei )에서 분리된 WSSV의 전장유전체 분석
이아름,공경희,김휘진,오명주,김도형,김종오,김위식,Lee, A-reum,Kong, Kyoung-Hui,Kim, Hwi-Jin,Oh, Myung-Joo,Kim, Do-Hyung,Kim, Jong-Oh,Kim, Wi-Sik 한국어병학회 2022 한국어병학회지 Vol.35 No.1
The full genome sequence of a Korean white spot syndrome virus (WSSV, isolate: WSSV-GoC18) is presented here. We obtained a total of 12,320,554 reads with 291,172 bases, 170 gene, and 170 coding DNA sequence, which were assembled in 1 contig. Phylogenetic analysis revealed that the WSSV-GoC18 was closely related to Chinese isolate (WSSV-PC) and distinctly different with previously reported a Korean isolate (WSSV K-LV1). The complete genome sequence of WSSV isolates will be of great help in molecular epidemiological studies, contributing to molecular diagnosis and disease prevention in shrimp aquaculture.
해수 사육 무지개송어(Oncorhynchus mykiss)의 Viral Hemorrhagic Septicemia Virus (VHSV) 모니터링
김위식,공경희,전영호,오명주,Kim, Wi-Sik,Kong, Kyoung-hui,Jeon, Young-Ho,Oh, Myung-Joo 한국수산과학회 2017 한국수산과학회지 Vol.50 No.5
Mariculture of rainbow trout Onchorhynchus mykiss has been initiated in or around olive flounder Paralichthys olivaceus farms, where viral hemorrhagic septicemia virus (VHSV) is often detected in some fish. In the present study, we investigated VHSV infection in seawater-reared rainbow trout because VHSV has never been detected in salmonids in Korea. A total of 104 adult fish were tested for the presence of VHSV by reverse transcription-polymerase chain reaction, followed by virus isolation with the fathead minnow caudal trunk cell line. Cytopathic effects were observed in two samples but the virus was identified as infectious hematopoietic necrosis virus. Thus, VHSV was not isolated from seawater-reared rainbow trout.
해수 사육 무지개송어(Oncorhynchus mykiss)의 위팽창증후군
김위식 ( Wi-sik Kim ),공경희 ( Kyoung-hui Kong ),오명주 ( Myung-joo Oh ) 한국어류학회 2017 韓國魚類學會誌 Vol.29 No.1
본 연구에서는 2014년 제주도에 위치한 양식장에서 해수로 사육 중인 무지개송어에서 만성질병으로 인해 약 10%의 누적 폐사가 발생하였다. 병어는 복부가 팽만되어 있었고 위가 심하게 팽창되어 있었다. 질병 검사 결과, 기생충, 세균, 바이러스는 검출되지 않았으나 팽창된 위에서 다수의 Candida sp.가 분리 되었다. Candida sp.를 사용하여 감염 실험을 실시한 결과, 대부분 개체에서 폐사가 발생하지 않았고 위 팽창 증상이 관찰되지 않아 Candida sp.는 본 질병의 원인 병원체가 아닌 것으로 사료되었다. 본 연구에서는 국내에서 처음으로 해수 사육 무지개송어에서 위팽창증후군이 발생하는 것을 확인하였다. About 10% mortality occurred in cultured rainbow trout Oncorhynchus mykiss at a marine farm in Jeju in 2014. Diseased fish showed markedly abdominal distension and distended stomach. Although parasites, bacteria or viruses were not isolated from diseased fish, numerous Candida sp. were isolated from distended stomach. In experimental infection with Candida sp., mortality was not observed in most fish and clinical sign of distended stomach was not observed in the fish. These results suggest that Candida sp. may not be the etiologic agent of stomach distension syndrome. This is the first report of stomach distension syndrome in seawater farmed rainbow trout in Korea.
단보 : 2014-2015년 남서해안 종묘장에서 생산된 넙치( Paralichthys olivaceus) 치어의 Kudoa septempunctata 감염실태 조사
김위식 ( Wi Sik Kim ),공경희 ( Kyoung Hui Kong ),정성주 ( Sung Ju Jung ),정명화 ( Myung Hwa Jung ),전찬혁 ( Chan Hyeok Jeon ),김정호 ( Jeong Ho Kim ),오명주 ( Myung Joo Oh ) 한국어병학회 2015 한국어병학회지 Vol.28 No.2
2014년 1월부터 2015년 5월까지 충청남도, 전라북도 및 전라남도에 위치한 6개지지역 (충청남도:태안, 보령, 전라북도: 고창, 전라남도: 무안, 영광, 완도) 총 11개소의 넙치 종묘장으로부터 채집한 넙치 치어 총 660마리 (132 pooling 시료)를 대상으로 Kudoa septempunctata의 감염현황을 조사하였다. 쿠도아 검사를 실시한 결과, 모든 넙치 종묘에서 PCR 음성 결과가 확인되었다. 이상의 결과로 쿠도아 진단 매뉴얼 방법에 의해 조사된 종묘장의 넙치에서는 K. septempunctata가 검출되지 않는 것으로 나타났다. A survey was conducted to investigate the infection of Kudoa septempunctata in 660 olive flounder (Paralichthys olivaceus) (132 pooling samples) cultured in 11 hatcheries in 6 different regions of Korea between 2014 and 2015. Polymerase chain reaction (PCR) results were negative for K. septempunctata for all samples. Based on the kudoa diagnostic manual, K. septempunctata was not detected in olive flounder hatcheries.
능성어(Epinephelus septemfasciatus) immunoglobulin M에 대한 단클론 항체 생산
김시우 ( Si-woo Kim ),김종오 ( Jong-oh Kim ),공경희 ( Kyoung-hui Kong ),오명주 ( Myung-joo Oh ),김위식 ( Wi-sik Kim ) 한국어병학회 2021 한국어병학회지 Vol.34 No.1
Immunoglobulin M (IgM) of sevenband grouper (Epinephelus septemfasciatus) was purified by mannan-binding protein (MBP) affinity column. The purified IgM had an apparent molecular weights of 76 (heavy chain) and 28 (light chain) kDa in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Eight hybridoma clones secreting monoclonal antibodies (mAbs) against sevenband grouper IgM were established. Antibody detection enzyme-linked immunosorbent assay (ELISA) with bovine serum albumin (BSA, antigen) and the 8 mAbs revealed that optical density (OD) values were clearly different between sera from BSA-immunization and non-immunization of sevenband grouper. These results suggest that the produced mAbs in this study are specifically reacted with IgM of sevenband grouper.
김위식 ( Wi-sik Kim ),정하나 ( Ha-na Jeong ),공경희 ( Kyoung-hui Kong ),김아라 ( Ah-ra Kim ),전영호 ( Young-ho Jeon ),오명주 ( Myung-joo Oh ) 한국어류학회 2016 韓國魚類學會誌 Vol.28 No.3
본 연구에서는 넙치 유래 VHSV (genotype IVa)에 의한 무지개송어의 영향을 조사하기 위하여 다양한 조건에서 감염 실험을 실시하였다. 3개의 다른 로트의 담수 무지개송어 치어에 VHSV (10<sup>6.3-7.3</sup> TCID<sub>50</sub>/fish)를 복강 주사한 결과, 15% 이하의 누적 폐사율이 관찰되었다. 무지개송어 (in vivo)를 사용하여 VHSV를 계대 배양한 후 감염 실험을 실시한 결과, VHSV를 5회 계대 배양하여도 독력의 변화는 관찰되지 않았다. 해수에 사육 중인 송어 (성어)에 넙치 유래 VHSV(10<sup>5.3</sup> TCID<sub>50</sub>/fish와 10<sup>6.3</sup> TCID<sub>50</sub>/fish)를 복강 주사한 결과, 폐사어는 관찰되지 않았고, 어체 내에서도 VHSV는 검출되지 않았다. 이상의 결과, 해수 송어 (성어)에서는 넙치 유래 VHSV의 병원성이 확인되지 않았으나 담수 무지개송어 치어에서는 낮은 독력을 나타내었다. Experimental infection of rainbow trout Oncorhynchus mykiss with viral hemorrhagic septicemia virus (VHSV, genotype IVa) from olive flounder Paralichtys olivaceus was examined. The cumulative mortalities of three different lot of rainbow trout fry challenged with VHSV (10<sup>6.3-7.3</sup> TCID<sub>50</sub>/ fish) were less than 15%. No difference of virulence was observed in experimental infection using 5 in vivo passaged VHSV and original VHSV. No mortality was observed in seawater-reared rainbow trout (adult) challenged with VHSV (10<sup>5.3-6.3</sup> TCID<sub>50</sub>/fish) and virus was not detected in the fish. We thus concluded that VHSV from olive flounder has low virulence to rainbow trout fry, but not pathogenic to seawater-rainbow trout (adult).