• Title/Summary/Keyword: viral neutralization

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Isolation and identification of infectious pancreatic necrosis virus form rainbow trout cultured in Korea (Isolation and Identification of Infectious Pancreatic Necrosis Virus from Rainbow Trout Cultured in Korea)

  • Lee, Jin-Hee;Heo, Gang-Joon;Lee, Chan-Hee
    • Journal of Microbiology
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    • v.34 no.2
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    • pp.124-130
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    • 1996
  • A survey was conducted to determine the prevalance of infectious pancreatic necrosis virus (IPNV) on fish farms in Korea and the epidemiology of IPNV infection in the farmed rainbow trout. In total, 43 pools of rainbow trout with apparent signs of viral infection from five provinces were obtained and analyzed. Evident cytopathic effects, including karyopycnosis and cell destruction, were observed in CHSE (chinook samlmon embyro)-214 cells infected with the virus isolates. Of these, ten viral isolates were assumed to be IPNV based on biophysical properties. RNA analysis revealed that the isolates contained two-segmented RNA genomes, further indicating that the viral isolates are IPNV. Antigenic comparison of the IPNV isolates identified three distinct serological groups separable by the cross-neutralization test. Of the ten IPNV isolates, six could be classified as strain DRT, two as strain Ab, and two as strain VR299. We were not able to isolate new strain of IPNV or any isolate serologically similar to the standard strain Sp.poraceae and families of the Agaricales, they are genetically more related to the Polyporaceae. These results are consistent with morphological characters observed in those mushrooms. However, it is premature to conclude taxonomic status Ganoderma species in the present study employing small sample size.

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Detection of viral-like particles in feces of Korean indigenous calves with diarrhea by negative staining (음성대조 염색법을 이용한 설사 한우송아지 분변내 바이러스양 입자 검색)

  • 임종수;강춘원;이태욱;김내영;정용운;강문일;한동운;최현성;이채용
    • Korean Journal of Veterinary Service
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    • v.22 no.1
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    • pp.43-52
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    • 1999
  • Presently, viral isolation in the diarrheal feces can be reached by many tools such as fluorescent antibody test(FA), negative contrast electron microscopy(NCEM), virus neutralization test, cell culture, and so on. The purpose of the study was to aimed at the establishment of simplified NCEM technique which can be efficiently applied for diarrheal feces and also the understanding on prevalence of viral-induced diarrhea in calves. One hundred fourty-seven korean indigenous calves with diarrhea were examined to their feces by the modified NCEM. Among them, 98(66.7%) were confirmed to have one or more viruses in feces. The viruses detected were identified as rotavirus(33.3%), coronavirus(16.3% ), togavirus(10.2%) and herpesvirus(0.7%). Ten cases of combined viral infection were consisted of 8 with rotavirus+coronavirus, one with rotavirus+togavlrus and one with rotavirus+herpesvirus. Dirrheal types could classified by yello-wish watery(44.9a ), blood-tinged(19.7% ), white watery(17.7% ) , brownish watery(14.3%), greenish watery(3.4%) diarrhea, respectively. Yellowish watery diarrhea(66cases) was frequently included rotavirus(31.8%), coronavirus(15.2%), and togavirus(13.6%), respectively. Consequently, these results suggest that the modified NCEM is reliable and efficient diagnostic tool for detection of viruses in the diarrheal feces and many calves rearing in Chonnam province have been exposed to some enteric viral agents mainly including rotavirus and coronavirus.

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Incidence of canine viral diseases and prevalence of virus neutralization antibodies of canine distemper virus, adenovirus type 2, parvovirus, and parainfluenza virus type 5 in Korean dogs

  • Dong-Kun Yang;Ha-Hyun Kim;Hye Jeong Lee;Young-Ju Cheong;Lee-Sang Hyeon;Minuk Kim;Bang-Hun Hyun
    • Korean Journal of Veterinary Research
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    • v.64 no.1
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    • pp.3.1-3.8
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    • 2024
  • Canine distemper virus (CDV), canine adenovirus type 2 (CAV-2), canine parvovirus (CPV), and canine parainfluenza virus 5 (CPIV-5) are the major viral pathogens in dogs. Despite the availability of vaccines for dogs against these 4 viral pathogens, investigations of antibodies against these pathogens have rarely been reported in South Korea. In this study, we investigated the recent incidence of viral diseases in dogs and conducted sero-surveillance for CDV, CAV-2, CPV, and CPIV-5 in Korean dogs. The most frequently diagnosed canine viral disease in Korean dog samples from 2000 to 2022 was CPV infection, which accounted for 48.7% (464/953) of the cases. A total of 400 dog serum samples collected between 2019 and 2022 were screened for the presence of virus-neutralizing antibodies against CDV, CAV-2, CPV, and CPIV-5. The overall seropositivity rates for CDV, CAV-2, CPV, and CPIV-5 were 83.8%, 77.8%, 99.3%, and 82.0%, respectively. The protection rate against CPV was the highest (98.3%) and that against CAV-2 was the lowest (44.8%) in dog sera. Male and female dogs showed no significant differences in seropositivity rates. CDV and CPIV-5 seropositivity increased with age in dogs, and the highest incidence and seropositivity rates of CPV indicated that Korean dogs have been continuously exposed to wild CPV, and that CPV is a pathogen that urgently requires attention among canine viral diseases.

Development of a Blocking ELISA for Measuring Rabies Virus-specific Antibodies in Animals

  • Yang, Dong-Kun;Kim, Ha-Hyun;Ryu, Jieun;Gee, Mi-ryun;Cho, In-Soo
    • Microbiology and Biotechnology Letters
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    • v.46 no.3
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    • pp.269-276
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    • 2018
  • Rabies virus (RABV)-specific antibodies in animals and humans are measured using standard methods such as fluorescent antibody virus neutralization (FAVN) tests and rapid fluorescent focus inhibition tests, which are based on cell culture systems. An alternative assay that is safe and easy to perform is required for rapid sero-surveillance following mass vaccination of animals. Two purified monoclonal antibodies (4G36 and B2H17) against RABV were selected as capture and detection antibodies, respectively. A genetically modified RABV, the ERAGS strain, was propagated and concentrated by polyethylene glycol precipitation. Optimal conditions for the RABV antigen, antibodies, and serum dilution for a blocking enzymelinked immune sorbent assay (B-ELISA) were established. We evaluated the sensitivity, specificity, and accuracy of the B-ELISA using serum samples from 138 dogs, 71 raccoon dogs, and 25 cats. The B-ELISA showed a diagnostic sensitivity of 95.8-96.3%, specificity of 91.3-100%, and accuracy of 96.0-97.2% compared to the FAVN test. These results suggest that the B-ELISA is useful for sero-surveillance of RABV in dogs, raccoon dogs, and cats.

Studies on Properties of Avian Reovirus Isolated in Korea (국내에서 분리한 닭 레오바이러스의 성상에 관한 연구)

  • 김성식;박병옥;김순재
    • Korean Journal of Veterinary Service
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    • v.15 no.1
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    • pp.67-80
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    • 1992
  • Avian reoviruses have been implicated in respiratory disease enteric conditions including Cloacal pasting in young thicks, pericarditis, hydropericardium, anaemia with swollen spleen and liver and petechiation of skeletal muscle and viral arthritis. This study was conducted to examine properties of reovirus field 3 strains isolated from affected broiler from several farms. An infectious agent was isolated from leg tendons and intestine of broiler with clinical tenosynovitis. The agent grew well on the chicken embryo kideny cells(CEK). One of them produced cytopathic effects(CPE) of round type and formed intranuclear inclusions, and the other was characterized by CPE of syncytical type and cytoplasmic inclusion. The properties and serological classification of field strains were examined by hemagglutin test, virus neutralization test, agar gel precipitin, electropherotype. They showed no hemagglutination reactions and not well neutralization and to possess common antigens detectable by AGP test. RNA electropherotype presented 10 segment band as the previous report. These data suggest that the field strains and standard strains (1133, 1733) may be the same group.

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Development of indirect ELISA for the detection of canine adenovirus type 2 antibodies in dog sera

  • Yang, Dong-Kun;Kim, Ha-Hyun;Lee, Siu;Oh, Dongryul;Yoo, Jae Young;Hyun, Bang-Hun
    • Journal of Veterinary Science
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    • v.21 no.4
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    • pp.63.1-63.9
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    • 2020
  • Background: Canine adenovirus type 2 (CAV-2) induces infectious laryngotracheitis in members of the family Canidae, including dogs. To date, no ELISA kits specific for CAV-2 antibody have been commercialized for dogs in Korea. Objectives: We aimed to develop new indirect enzyme-linked immunosorbent assay (I-ELISA) to perform rapid, accurate serological surveys of CAV-2 in dog serum samples. Methods: In total, 165 serum samples were collected from dogs residing in Chungbuk and Gyeongbuk provinces between 2016 and 2018. The Korean CAV-2, named the APQA1701-40P strain, was propagated in Madin-Darby canine kidney cells and purified in an anion-exchange chromatography column for use as an antigen for I-ELISA. The virus-neutralizing antibody titers of CAV-2 in the dog sera were measured by virus neutralization (VN) test. Results: We compared the results obtained between the VN and new I-ELISA tests. The sensitivity, specificity, and accuracy of new I-ELISA were 98.6%, 86.4% and 97.0% compared with VN test, respectively. New I-ELISA was significantly correlated with VN (r = 0.91). Conclusions: These results indicate that new I-ELISA is useful for sero-surveillance of CAV-2 in dog serum.

Diagnosis of bovine virus diarrhea-mucosal disease in breeding Holstein cows with diarrhea and vesicles (설사 및 수포발생 육성유우에서 바이러스성 설사.점막병의 진단)

  • 이성효;한수철;이종오;윤여백;송희종;채효석
    • Korean Journal of Veterinary Service
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    • v.20 no.4
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    • pp.331-338
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    • 1997
  • The purpose of this study was to establish early diagnosis for bovine virus diarrhea-mucosal disease(BVD-MD) Two Holstein among 22 breeding cows were shown ulcer in the mouth and watery diarrhea. Diarrheal feces and ulcerous lesion of the mouth from 2 cows were sampled for detection of viral antigen. BVD virus was isolated by inoculation of the samples to MDBK cells, and the cytopathic effects were observed in cultured MDBK cells which inoculated with virus isolates from the feces. Viral antigens were detected in the feces and ulceruous lesion by immunogold staining. The serum neutralization titers were shown 1 : 64 or greater in 8 blood samples by using BVD virus (NADL strain). By the RT-PCR, using reverse primer 5'-ACTCCATGTGCCATGTACAG-3', forward primer 5'-ACTCCATGTGCCATGTACAG-3', 285 base pair band specific to BVD virus was detected. In conclusions, the results of above tests which executed using the diarrheal feces and ulcerous lesion of the mouth and the isolates were conformed as BVD virus.

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Immunocytochemical identification of Aujeszky's disease viral antigens in nasal and circulating white blood cells from living pigs for early diagnosis (오제스키병의 생체 조기진단을 위한 돼지 비세포 및 순환백혈구내 면역세포화학적 바이러스 항원동정)

  • Moon, Oun-kyong;Cho, Hee-tack;Kim, Soon-bok;Sur, Jung-hyang
    • Korean Journal of Veterinary Research
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    • v.33 no.2
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    • pp.241-247
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    • 1993
  • Forty day old piglets were intranasally inoculated with 2ml of Aujeszky's disease virus (NYJ-1-87 strain, $10^{7.0}$ $TCID_{50/0.2}ml$), and the viral antigens were detected in nasal and circulating white blood cells for 20 days after inoculation by immunocytochemical method. Antibody titers in the blood were also detected by neutralizing test and Aujeszky's disease serodiagnostic kit(Choong Ang) in this periods. 1. Viral antigens were detected by the immunocytochemical technigue, and positive reactions were observated in nasal cells from the 2nd to the l0th days after inoculation and circulated white blood cells from the 4th to the 12th days after inoculation. 2. In neutralization test antibodies levels showed titers of 2 on the 8th day, 8 on the l5th day, 16 on the 18th day and 32 on the 20th day after inoculation. In serodiagnostic kit test positive reactions were observed after the 15th day after inoculation.

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Monoclonal antibodies against structural proteins of bovine viral diarrhea virus (소 설사병 바이러스 구조단백에 대한 단크론항체 성상에 대한 연구)

  • Kweon, Chang-hee;Zee, Yuan Chun;Woo, Hee-jong
    • Korean Journal of Veterinary Research
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    • v.32 no.1
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    • pp.83-90
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    • 1992
  • Monoclonal antibodies against structural proteins of bovine viral diarrhea virus(BVDV) were derived by classical hybridoma techniques. These antibodies were characterized by serum neutralization, immunoblotting and immunoprecipitation. The neutralizing monoclonal antibody reacted with the 56kd to 54kd(M.W.) viral protein in western blotting and immunoprecipitation analysis. Although there was no neutralizing activity, another monoclanal antibody reacted with the 45kd protein by immunoprecipitation and with both the 45kd and 36kd proteins in immunoblotting analysis. respectively. Densitometer scanning of purified BVDV and the immunopreipitation of whole virus particles with neutralizing monoclonal antibody revealed the presence of more than twelve viral polypeptides. Although no possible precursor form of protein was identified with the neutralizing monoclonal antibody. the presence of intact virion was detected in the infected cell supernatant immediatelty after pulse labeling, indicating rapid translational processing as well as packaging of the virus. The partial peptide mapping of 45kd and 36kd proteins with Staphylococcus aureus V 8 protease showed that these two proteins are related.

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Experimental Infection for the Neutralization of White Spot Syndrome Virus (WSSV) in Wild Captured Sand Shrimp, Crangon affinis (자연산 자주새우(Crangon affinis)에서 흰반점바이러스(WSSV)의 neutralization을 위한 인위감염(experimental infection))

  • Gong, Soo-Jung;Kim, Yeong-Jin;Choi, Mi-Ran;Kim, Sung-Koo
    • Journal of Life Science
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    • v.20 no.9
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    • pp.1294-1298
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    • 2010
  • White spot syndrome virus (WSSV) is one of the most virulent viral agents threatening the penaeid shrimp culture industry. This study was carried out to evaluate the susceptibility of the sand shrimp, Crangon affinis, to WSSV as an alternative experimental model. WSSV caused 100% mortality in C. affinis within 7 days after experimental infection by immersion. Based on challenge studies, it was confirmed that C. affinis could be a potential host in WSSV transmission. Also, the neutralization of WSSV was carried out using an antiserum raised against recombinant envelop protein rVP466 to evaluate the WSSV infection mechanism. A constant amount of WSSV (at $1{\times}10^4$ diluted stocks) was incubated with various amounts of antiserum and then mixed to 20 l reservoir for the immersion challenge of C. affinis for neutralization. At 5 days post challenge, the shrimp in the positive control immersed in the immersion reservoir containing WSSV stock showed 100% mortality. The shrimps challenged with the 3 different mixtures of WSSV and rVP466 antiserum (1:0.1, 1:0.5 and 1:1) showed 100%, 68.8% and 68.8% mortality at 14 days post challenge, respectively. These results indicated that the antiserum raised against rVP466 could block WSSV infection in C. affinis. Therefore, this study confirmed that C. affinis can be naturally infected by WSSV as another potential host and that C. affinis can be used as an alternative experimental animal instead of penaeid shrimps.