• 한국수의병리학회지
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• 제7권1호
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• pp.1-4
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• 2003

Malignant catarrhal fever (MCF) is a systemic disease of ruminants caused by a gamma herpesvirus, ovine herpesvirus 2 (OvHV-2). Dot blot hybridization (DBH) protocols for detecting and differentiating this MCF virus were developed. OvHV-2 specific primer pairs, 556/555, were used for the amplification of target DNA. Then, the amplified DNA was labeled with incorporation of digoxigenin (DIG). The Dig-labeled probe was able to detect and differentiate specifically OvHV-2 DNA. This DBH technique can be applied to confirm the presence of MCF virus on clinical samples and to differentiate specifically between OvHV-2 infection and other viral infections.

• 한국수의병리학회지
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• 제7권1호
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• pp.5-9
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• 2003

Malignant catarrhal fever (MCF) is a systemic disease of ruminants caused by a gamma herpesvirus, ovine herpesvirus 2 (OvHV-2). Four 1-year old goats (Capra hircus), which were infected with MCF virus, OvHV-2, by being housed together with MCF virus-infected seep, were referred with a I-month history of chronic dermatitis. On the other hand, MCF virus-negative goats, which were isolated for negative control, had not those kinds of skin problems. Examination of the affected goats revealed generalized alopecia, patchy erythema, and superficial erosions with histologic evidence of mural folliculitis. Fungal culture tests and external parasite tests with the scraping skin samples were negative. However, polymerase chain reaction revealed the existence of MCF virus DNAs in the lesion. These results suggested that MCF virus may induce mural folliculitis and alopecia in goat.

• Journal of Veterinary Science
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• 제22권4호
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• pp.51.1-51.10
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• 2021

Background: Malignant catarrhal fever (MCF) is a highly fatal lymphoproliferative disease of cattle, deer, bison, water buffalo, and pigs caused by the gamma-herpesviruses alcelaphine herpesvirus-1 (AlHV-1) and ovine herpesvirus-2 (OvHV-2). Objectives: This study aimed to determine the prevalence of OvHV-2 in sheep, goats, cattle, and buffalo in Rawalpindi and Islamabad, Pakistan, by applying molecular and phylogenetic methods. Methods: Blood samples were aspirated from sheep (n = 54), goat (n = 50), cattle (n = 46) and buffalo (n= 50) at a slaughterhouse and several farms. The samples were subjected to heminested polymerase chain reaction (PCR), followed by sequencing and phylogenetic analysis of the OvHV-2 POL gene and the OvHV-2 ORF75 tegument protein gene. Results: The highest percentage of MCF positive samples was in sheep (13%), whereas goat, cattle, and buffalo had lower positive percentages, 11%, 9%, and 6.5%, respectively. Four OvHV-2-positive PCR products obtained from sheep samples were sequenced. The sequences obtained were submitted to the NCBI GenBank database (MK852173 for the POL gene; MK840962, MK852171, and MK852172 for the ORF75 tegument protein gene). Phylogenetic analysis revealed a close similarity of study sequences with those of worldwide samples. Conclusions: This study is the first cross-sectional study on the prevalence and molecular detection of OvHV-2 in apparently healthy cattle and buffalo that could be carrying OvHV-2 acquired from OvHV-2-positive sheep and goats. The results indicate that OvHV-2 is circulating in Pakistan. Further studies are needed to characterize OvHV-2 and elucidate further its prevalence.

• 대한수의학회지
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• 제43권3호
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• pp.471-476
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• 2003

Malignant catarrhal fever (MCF) is a systemic disease of ruminants caused by ovine herpesvirus 2 (OvHV-2). OvHV-2 is a gamma herpesvirus, which induces frequent latent infection and often difficult to detect its antigens and even specific nucleic acids because of its low viral copies in the infected tissues. Histopathology, serology and polymerase chain reaction (PCR) were compared for the diagnosis of MCF using 10 bison infected with OvHV-2. Histopathological diagnosis was performed using the criteria which was based upon the pathognomic lesions. Serological diagnosis was conducted using its serum with competitive ELISA for the detection of antibodies of OvHV-2. Also, the nest PCR was performed with peripheral blood leukocytes for the detection of OvHV-2-specific DNAs. Primers 556 and 775 were used for the primary amplification, and primers 556 and 555 were used for the secondary amplification. As the results, positive cases were 6 by histopahology, 9 by serology and 10 by PCR. As comparing with other diagnostic methods, PCR was found to be more sensitive than histopathology and serology. The recent development of molecular diagnostic assays has provided powerful tools for investigating how viruses survive in nature. Development of PCR specific for viruses has dramatically improved the accuracy of diagnosis of viruses in clinically infected animals. Furthermore, amplification of viral genomic material by nest PCR represents the most sensitive method for the detection of viruses and might be detected successfully even though very low viral DNA copies. So, it could be used as the first choice for the detection of viral DNAs with low copies such as the status of latent infection. However, it has also some limitation of application like as false negative results by PCR inhibitors and false positive results by contamination. The results of this study suggest that the use of molecular biological methods like PCR may increase the accuracy for the diagnosis of infectious diseases. However, in diagnostic laboratory, it is recommended that PCR assay must be conducted with other diagnostic methods for more reliable diagnosis.

• 한국수의병리학회지
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• 제7권1호
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• pp.11-15
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• 2003

Antigen retrieval (AR) techniques were widely used to recover the antigenicity from the fixed tissues, which were guided by the philosophy of rendering immunohistochemistry (IHC) applicable to routine formalin-fixed, paraffin-embedded tissues for wide application of IHC in research and clinical filed for morphological observation like as anatomy, histology and pathology. Protease antigen recovery (PAR) is an AR technique, which is obtained the antigen retrieve by using enzyme digestion, and commonly used in IHC field. However, during the IHC for the detection of ovine herpesvirus 2 (OvHV-2) antigen, we noted lymphocyte-like cells-specific staining in the infiltrated cells into various organs like as liver and kidney, which was also shown in the IHC tissues with isotype control. However, those signals were not observed in the tissues conducted with in situ hybridization. Therefore, we analyzed the specificity of the IHC detection results. We found that PAR may induce false-positive result during IHC in lymphocyte-like cells, which were infiltrated mainly around vessels and in interstitial tissues. Through the Phenotyping, we realized that those false-positive cells were B-cell-related cells. These results suggest that PAR, a AR using protease, may induce non-specific false-positive reactions during IHC.