• 한국수의병리학회:학술대회논문집
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• 한국수의병리학회 2002년도 추계학술대회초록집
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• pp.142-142
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• 2002

돼지의 주요 장염 유발 바이러스인 돼지 전염성 위장염 바이러스 (transmissible gastroenteritis virus; TGEV), 돼지 유행성 설사증 바이러스 (porcine epidemic diarrhea virus; PEDV), 돼지 칼리시 바이러스 (porcine enteric calicivirus; PECV), 돼지 로타바이러스 A 형과 C 형 (porcine rotavirus; PRY, group A and C)을 동시에 감별 진단 할 수 있는 신속하고 정확한 oligonucleotide microarray 진단법을 개발하였다. 이 진단법은 유리슬라이드에 각각의 바이러스에 특이적인 부위에서 제작된 oligonucleotide probe를 찍은 DNA chip을 제작하여 여기에 각각의 바이러스를 역전사하고 cy5-dCTP를 포함한 multiplex PCR을 수행한 다음 hybridization 하였다. 이후 hybridization 결과는 fluorescence scanner를 이용하여 확인하였다. 이 새로운 microarray system은 RT-PCR과 같은 기존의 진단방법보다 소량의 바이러스를 민감하게 검사할 수 있을 뿐 아니라 hybridization을 통해 검사결과의 정확성을 확인할 수 있었다. 따라서 본 연구에서 개발한 microarray system은 돼지의 설사 유발 바이러스를 진단하는데 매우 유용한 진단 방법임을 확인하였다.

• Journal of Microbiology and Biotechnology
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• 제30권4호
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• pp.515-525
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• 2020

Interferon (IFN)-λ plays an essential role in mucosal cells which exhibit strong antiviral activity. Lactobacillus plantarum (L. plantarum) has substantial application potential in the food and medical industries because of its probiotic properties. Alphacoronaviruses, especially porcine epidemic diarrhea virus (PEDV) and transmissible gastroenteritis virus (TGEV), cause high morbidity and mortality in piglets resulting in economic loss. Co-infection by these two viruses is becoming increasingly frequent. Therefore, it is particularly important to develop a new drug to prevent diarrhea infected with mixed viruses in piglets. In this study, we first constructed an anchored expression vector with CWA (C-terminal cell wall anchor) on L. plantarum. Second, we constructed two recombinant L. plantarum strains that anchored IFN-λ3 via pgsA (N-terminal transmembrane anchor) and CWA. Third, we demonstrated that both recombinant strains possess strong antiviral effects against coronavirus infection in the intestinal porcine epithelial cell line J2 (IPEC-J2). However, recombinant L. plantarum with the CWA anchor exhibited a more powerful antiviral effect than recombinant L. plantarum with pgsA. Consistent with this finding, Lb.plantarum-pSIP-409-IFN-λ3-CWA enhanced the expression levels of IFN-stimulated genes (ISGs) (ISG15, OASL, and Mx1) in IPEC-J2 cells more than did recombinant Lb.plantarum-pSIP-409-pgsA'-IFN-λ3. Our study verifies that recombinant L. plantarum inhibits PEDV and TGEV infection in IPEC-J2 cells, which may offer great potential for use as a novel oral antiviral agent in therapeutic applications for combating porcine epidemic diarrhea and transmissible gastroenteritis. This study is the first to show that recombinant L. plantarum suppresses PEDV and TGEV infection of IPEC-J2 cells.

• 대한수의학회지
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• 제43권2호
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• pp.219-230
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• 2003

Porcine epidemic diarrhea virus(PED), a member of Coronaviridea, is the etiological agent of enteropathogenic diarrhea in swine. The purpose of this study was to investigate genetic characteristic of PEDV isolated in Korea. Nucleocapsid(N) gene and membrane (M) gene of recent Korean PEDV strains isolated in 2001 were amplified, cloned, sequenced and analyzed. N gene of seven Korean PEDV field isolates bad 94.5% to 99.4% nucleotide and 92.4% to 99.4% amino acid sequence homology each other. Nucleotide and amino acid sequences of Korean field PEDVs were different from published foreign PEDVs, showing 95.1% to 98.0% nucleotide and 93.5% to 97.6% amino acid sequence homology. By phylogenetic tree analysis on based nucleotide sequences, PEDVs were clustered into four groups. By phylogenetic tree analysis based on amino acid sequences. PEDVs were clustered into five groups. M gene of our Korean PEDV field isolates had 99.6% to 100% nucleotide and 98.7% to 100% amino acid sequence homology each other. Nuclotide and amino acid sequences of Korean field PEDVs were different from published foreign PEDVs, showing 98.5% to 98.8% nucleotide and 97.3% to 97.8% amino acid sequence homology. By phylogenetic tree analysis based on nucleotide and amino acid sequences, PEDVs were clustered into two groups which were Korean PEDV isolate group and foreign PEDV isolate group.

• 한국동물위생학회지
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• 제39권4호
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• pp.259-266
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• 2016

Porcine epidemic diarrhea virus (PEDV) causes an acute and lethal watery diarrhea in piglets that is great economic losses to the swine country worldwide. The spike (S) glycoprotein is an important determinant for PEDV biological properties. In the present study, we determined the full-length S gene sequences of five Chung-nam PEDV field isolates collected in 2016. The S gene was amplified by RT-PCR, purificated, sequenced, analyzed and then compared with published sequences of other PEDV strains. 5 field strains share 98.5%~99.9% homologies with each other at the nucleotide sequence level and 96.7%~99.9% homologies with each other at the amino acids sequence level. Most field strains have nucleotide insertions, deletions and mutation regions, and show lower homologies (93.1~93.8%) with classical and vaccine strains, however higher homologies (99.1%~99.5%) with US PEDV isolates in 2013. By phylogenetic tree analysis based on nucleotide sequence, five PEDV field isolates were clustered into Genogroup 2b but differ genetically from the vaccine strains (SM-98 and DR-13).

• Journal of Plant Biotechnology
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• 제32권4호
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• pp.263-268
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• 2005

Porcine epidemic diarrhea virus (PEDV)는 돼지의 급성장염을 유발하여 설사 증상을 일으키는 바이러스이다. 본 연구에서는 고구마 저장뿌리 고발현 sporamin 프로모터 및 CaMV 35S promoter를 이용하여 PEDV 항원단백질을 생산하는 고구마 식물체를 개발하고자 하였다. 형질전환 벡터를 제작하기 위하여 PEDV에서 항원성이 알려진 스파이크 단백질의 일부분을 암호화하는 유전자를 PCR로 합성하였다. 고구마 [Ipomoea batatas (L.) Lam] 율미 품종의 배발생 캘러스를 재료로 하여 Agrobacterium tumefaciens을 매개로 형질전환하였다. 선발배지 (MS medium, 1 mg/L 2,4-D, 100 mg/L kanamycin, and 400 mg/L claforan)에서 배발생 캘러스를 3주 간격으로 4개월 동안 계대배양하여 카나마이신 저항성 캘러스를 선발하였다. 선발된 배발생 캘러스를 호르몬을 제거한 배지로 옮겨 체세포배를 유도하였으며 이후 shoot과 뿌리가 형성되었다. 재분화된 소식물체를 대상으로 PCR 분석으로 카나마이신 저항성 재분화 개체의 50% 이상이 도입 유전자를 가지고 있었으며 이들 식물체를 대상으로 Southern blot 분석하여 PEDV 유전자가 고구마 식물체의 게놈으로 안정적으로 도입되었음을 확인하였다. 형질전환 식물체에서 도입유전자의 발현을 RT-PCR로 분석한 결과 PEDV의 spike 유전자가 높은 수준으로 발현하였다.

• 한국수의병리학회:학술대회논문집
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• 한국수의병리학회 2003년도 추계학술대회초록집
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• pp.32-32
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• 2003

Intestinal infections are common in growing pigs and can be caused by multiple pathogens, environmental and management factors [1]. Among the most important viruses in swine enteritis are porcine epidemic diarrhea virus (PEDV), transmissible gastroenteritis virus (TGEV), porcine enteric calicivirus (PECV), porcine group A rotavirus (PRV gp A) and bacteria are Escherichia coli and Salmonella spp. and protozoa is Isospora suis [1]. The DNA chip system can serve as a powerful tool that can be utilized for simultaneous detection of specific pathogenic bacteria strains and viruses [2,3]. The combination of PCR and DNA chip technology will provide a novel method for the detection of porcine enteric pathogens thus revolutionize the diagnosis and management of the disease. The aim of this study is to develop DNA chip system for the rapid and reliable detection of five major porcine enteric pathogens based on oligonucleotide DNA chip hybridization. (omitted)

• 한국식물생명공학회:학술대회논문집
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• 한국식물생명공학회 2002년도 춘계학술대회
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• pp.163-163
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• 2002

• 한국식물생명공학회:학술대회논문집
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• 한국식물생명공학회 2005년도 추계학술대회 및 한일 식물생명공학 심포지엄
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• pp.419-419
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• 2005

• 한국식물생명공학회:학술대회논문집
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• 한국식물생명공학회 2005년도 추계학술대회 및 한일 식물생명공학 심포지엄
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• pp.146-146
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• 2005

• 한국동물위생학회지
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• 제43권4호
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• pp.237-244
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• 2020

Porcine transmissible gastroenteritis (TGE) has been a significant cause of economic losses in pig farming industry since 1950s. Although transmissible gastroenteritis virus (TGEV) has declined in recent years, it should not be excluded because of its characteristics; the frequency of gene mutation, the mortality in piglets, and the possibility for sudden incidence. Therefore, the herd-level monitoring of the virus is important to prevent further circulation of TGE. The aim of this study is to develop a large-scale sandwich enzyme-linked immunosorbent assay (ELISA) with high specificity to rapidly detect TGEV in feces by using monoclonal antibodies (Mabs). The TGEV specific Mabs were produced in hybridoma cells. Among the Mabs belonged to the IgG class developed by this study, the final selected 8H6, 1B7, 4G3, and 1F8 were identified to have the neutralization ability against TGEV. The sandwich ELISA was established using 8H6 as a reporter antibody and 1B7 and the reported 5C8 as a capture antibody. The developed sandwich ELISA was able to distinguish TGEV from other pathogenic diarrheal agents (porcine rotavirus, porcine reovirus, porcine epidemic diarrhea virus (PEDV), E. coli, and C. perfringens) in tissue culture as well as fecal samples. And the detection rate of TGEV in feces was 80% compared with RT-PCR. The results suggested that the developed sandwich ELISA may be useful in the herd-level monitoring for effective preventive measures due to the early diagnosis of TGEV using a large amount of samples.