• Korean Journal of Veterinary Research
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• v.28 no.2
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• pp.365-369
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• 1988

The present study was done to demonstrate ADV antigens in frozen and paraffin sections from ADV-infected pigs and cell cultures by using of the IGS method. Tissue specimens from 3 young pigs infected with ADV-phylaxia strain and of 2 healthy pigs were used. Fibroblastic cells originated from pig brain and BHK cells were grown and confluent monolayers were infected with the virus. Two monoclonal antibodies and a specific hyperimmune serum to ADV were used as the source of primary antibodies for both the IGS and immunoperoxidase methods. Application of the IGS method yielded a black fine granular reaction in positive areas, and the results were superior to those obtained using the immunoperoxidase technique for all cases tested. The IGS method might be useful in the detection of various viral antigens in tissue sections.

• Journal of the korean veterinary medical association
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• v.24 no.3
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• pp.163-171
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• 1988

A swine farm located in Namyangju, Kyunggi-do was damaged by increased loss due to stillbirth, abortion and high mortality of suckling pig during October to December 1987. Three piglets of the farm, that were diagnosed clinico-pathologically to be affecte

• Korean Journal of Veterinary Research
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• v.32 no.3
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• pp.365-368
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• 1992

1988년부터 1990년 6월까지 전국의 양돈장에서 수집된 돼지 유사산 태아 74복에서 바이러스성 원인체 분리 및 혈청학적 진단을 수행하였던 바 다음과 같은 결과를 얻었다. 공시한 74복의 유사산 태아중 44복의 태아 흉강액에서 면역 globulin이 검출되어 전염성 질병감염에 의한 유사산으로 추정되었다. 이중 37%가 바이러스성 유사산으로 나타났으며 유사산의 원인체별 분포를 살펴보면 돼지 파보바이러스가 21%로 가장 높았으며, 뇌심근염 바이러스가 11%, 일본뇌염 바이러스가 9% 등의 순으로 나타났다. 한편 돼지 콜레라바이러스 및 오제스키병 바이러스에 의한 유사산이 각각 1건씩 검출되었으며 동일 유사산 태아에서 2가지 병원체가 중복감염된 예도 관찰되었다.

• Korean Journal of Veterinary Research
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• v.28 no.1
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• pp.99-103
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• 1988

The first outbreak of Aujeszky's disease(AD) was identified from piggery located at the southern part of Korea in July, 1987. This piggery suffered from a significant economic loss caused by unexpected piglet mortality and reproductive failure. Etiologic viral agents were isolated from tonsil and spleen of the infected piglets, and the isolates produced a typical cytopathic effects of herpesvirus with giant cell formation when inoculated in many different cells. Subsequently the field isolates were characterized as suid herpesvirus I by cross-neutralization test and indirect fluorescence assay utilizing specific monoclonal antibody, and were proved to be a pathogenic strain of AD virus(ADV).

• Korean Journal of Veterinary Pathology
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• v.1 no.2
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• pp.119-124
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• 1997

The objective of this study was to develop digoxigenin (DIG)-labeled in situ hybridization (ISH) test for diagnosis of Aujeszky's Disease(AD) in infected organs. Specific DNA with well conserved gene sequences encoding gp50 antigen in AD virus (ADV) was obtained by Polymerase Chain Reaction (PCR) method. A pair of oligonucleotide primers used in PCR allowed amplification of a 217 bp sequence from the gp50 ADV gene. The DNA was then labeled with DIG by primer labeling method for use as probe in ISH test to detect ADV nucleic acids in various tissue. Positive hybridization was demonstrated by dark pigmentation in nuclei and cytoplasm of ADV infected cells particularly in brain tonsillar crypt epithelium and pulmonary alveolar cells. This result suggests that ISH is a valuable sensitive and rapid diagnostic test for AD.

• 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.

• Korean Journal of Veterinary Research
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• v.34 no.2
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• pp.327-333
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• 1994

The purpose of this study was to establish a rapid diagnostic method detecting Aujeszky's disease virus (ADV) DNA in the cultured cell monolayers (PK-15) and tissue sections of ADV(NYJ-1-87)-infected rats and pigs by in situ hybridization(ISH). Detection of specific ADV-DNA in infected cells was conducted by radiolabeled ISH method using $^{32}P-labeled $ DNA probe (BamH1 7 fragment) which contains a 6.3 Kb ADV-DNA insert. Where ADV-DNA was detected by radiolabeled ISH, the deposition of black photographic grains occurred in the nuclei and the cytoplasms of ADV-infected cells. Positive hybridization signal was often observed in the spinal trigerminal nucleus of the pons, the nucleus of the trigerminal ganglion neuron and the epithelial cells of tonsillar crypts. The results suggested that ISH is considered as a highly sensitive and reliable tool for confirmative diagnosis of this viral disease.

• Korean Journal of Veterinary Research
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• v.43 no.2
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• pp.239-246
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• 2003

Polymerase chain reaction (PCR) was evaluated for the early detection of Aujeszky's disease virus (ADV) DNA from virus-infected cell cultures. For the purposes, the Korean ADV NYJ1-87 was propagated in swine kidney (SK) cells and subjected to the amplification of DNA (217 bp) by PCR using sense and antisense primers specific to gp50 gene of the ADV. In detection of cell-associated viral DNA, reliable PCR conditions were determined as 30 cycles of reaction consisting 1 minute each of denaturation at $94^{\circ}C$, annealing at $55^{\circ}C$ and polymerization at $72^{\circ}C$. The PCR encountered best results with reagent mixtures of $50{\mu}l$ containing $200{\mu}M$ dNTPs, $0.2{\mu}M$ each sense and antisense primers, 1 mM $MgCl_2$ and 10% (v/v) template DNA in the final concentrations. ADV-specific DNAs were detected as early as 6, 6, and 9 hours post-infection, respectively, from lysates of the SK cells infected with ADV of $10^3$, $10^2$ and $10^1\;TCID_{50}/ml$ by this condition. In culture supernatant, the DNAs were detected from ADV of as low infectivity as $10^ {-3}\;TCID_{50}/ml$ by the reduced reagent concentrations and 30 cycles of 1 minute each of denaturation at $94^{\circ}C$ and annealing at $55^{\circ}C$, and 2 minutes of polymerization at $72^{\circ}C$. The lowest amount of detectable ADV DNA was 1 fg. In conclusion, the PCR condition established in the present study was recognized as a feasible alternative to time-consuming procedures in isolation and characterization of the virus.

• Korean Journal of Veterinary Research
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• v.37 no.4
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• pp.765-772
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• 1997

제주지역 돼지에서 각종 전염성 질병 원인체에 대한 항체를 조사하여 그간 전염성 병원체에 대한 역학조사가 미진하였던 부분을 보완하여 질병의 분포를 파악하고자 1995년부터 1996년에 걸쳐 제주도 전역에서 돼지의 혈청을 채취하여 각종 병원체에 대한 항체 분포율을 조사하였다. 본 연구에서 검사한 돼지 혈청 시료에서는 돼지 오제스키병 바이러스에 대한 항체는 전혀 검출되지 않았다. 돼지 콜레라바이러스에 대한 항체는 기대 수준 이하로 낮아 백신접종이 원활히 수행되고 있지 않음을 시사하였으며 특히 농장에 따라 항체 보유돈과 항체 음성돈이 혼재하는 농장과 항체가 전혀 검출되지 않는 농장 등 돼지 콜레라 방역의 사각지대가 존재할 가능성이 있음을 보여주었다. 유 사산 원인체인 돼지 파보바이러스 및 뇌심근염에 대한 항체가가 다양하게 나타나 일부 문제가 있을 것으로 사료되었다. 돼지 생식기호흡기증후군(PRRS) 바이러스에 대한 항체 분포율은 내륙 보다 다소 낮게 나타났고, 돼지 influenza virus, 위축성 비염, 흉막 폐염 등 각종 세균성 질환에 대한 항체수준도 다양하게 나타났다. 본 혈청학적인 연구결과는 제주지역에서의 양돈방역 정책수립 및 질병방제의 기초자료로 유용하게 이용될 것으로 사료된다.

• Korean Journal of Veterinary Pathology
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• v.2 no.2
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• pp.117-125
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• 1998

Pathological studies were performed on the five piglets experimentally infected with Aujeszky's disease virus(pseudorabies), NYJ isolate, isolated from the naturally infected pigs in Korea: two piglets were inoculated intramuscularly, two piglets intranasally, and one piglet subcutaneously at the dose of 1$m\ell$ per animal with the 105.5 $TCID_50$/0.1ml titer. Clinical signs included dyspnea, high fever(＞$41^{\circ}C$), anorexia, vomiting, diarrhea or constipation, ataxia, circling movement, posterior paralysis, intermittent convulsion, and coma followed by death although some variations by age and inoculated routes were observed. Gross features included multiple necrotic foci in the liver, congestion and hemorrhage in the lymph nodes and spleen, petechial hemorrhage in the kidney, hemorrhagic pneumonia, marked meningeal congestion, severe sub meningeal hemorrhage in the spinal cord, excessive cerebrospinal fluid retention, and muscular necrosis at the inoculated area. Microscopically, non suppurative meningoencephalitis with gliosis and perivascular cuffing in CNS, ganglioneuritis, necrohemorrhagic splenitis, necrotic hepatitis, tonsillitis and rhinitis, hemorrhagic or interstitial pneumonia, and non-suppurative myositis in the injected area were observed. Eosinophilic intranuclear inclusion bodies were found in a variety of tissues the including the liver, kidney, adrenal gland, spleen, lymph nodes, tonsil, and lung. Ultrastructurally, virus particles were confirmed in nucleus and cytoplasms of pneumocytes around the necrotic areas.