• Journal of fish pathology
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• v.26 no.3
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• pp.163-171
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• 2013

Polymerase chain reaction (PCR) is the most rapid and widely used method to detect viral pathogens. However, this method does not provide histopathologic nature of the virus. In situ hybridization (ISH) with oligonucleotide probes is attractive because it is a rapid method for detection and identification of viral pathogens at sites of tissue infection. In order to understand the histopathologic characterictics of Red sea bream iridovirus (RSIV), viral-hemorrhagic septicemia (VHS) virus and viral nervous necrosis (VNN) virus to cultured olive flounder, we her applied ISH method to various kinds of olive flounder tissues with PCR-positive for these three viruses. We found that these viruses showed different tissue tropism and were detected from different cell types. Our results suggest that ISH is useful not only in rapid detection of viral pathogens but also in understanding the histopathologic characters of specific viral pathogens.

• Korean Journal of Environmental Biology
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• v.18 no.1
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• pp.53-61
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• 2000

In order to characterize the genetic diversity of bacterial community in groundwater, samples were collected from used for drinking water and polluted with heavy metal wastewater in Seoul city and natural cave of Kangwondo. The DNA was amplified with 165 rDNA-based primers by use of the PCR, and then analysed ARDRA (amplified ribosomal DNA restriction analysis). Restriction endonuclease analysis patterns of amplified 165 rDNA in drinking water and wastewater relatively showed high genetic diversity in situ and drinking groundwater. The number of DNA fragments varied with in situ and drinking water. This method of ARDRA of bacterial communities in groundwater could be used for a quick assessment of genotypic changes between different locations reflecting different environmental conditions and the diversity reflected pollution of groundwater (natural cave water>drinking water>waste water, as in order of grade). [Genetic diversity, Groundwater, 165 rDNA, PCR, ARDRA].

• Korean Journal of Veterinary Service
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• v.40 no.3
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• pp.161-168
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• 2017

Caprine arthritis encephalitis (CAE) virus is a causative agent of caprine arthritis-encephalitis. In our previous study we reported a prevalence of CAE. In this study, we described the further detailed pathological features of CAE and examined the detection of virus by in situ hybridization (ISH). Histopathologically, interstitial pneumonia and bronchopneumonia in lung, focal inflammation in mammary glands, perivascular cuffing in brain, arthritis, and focal necrosis, mild steatosis, inflammatory cell infiltration of liver were noted. CAEV proviral-DNA was identified by nested polymerase chain reaction (PCR) in blood cells, brain, synovial fluid, and lymph node. Confirmation by nested PCR involved amplification of a 296 bp ($1^{st}$ PCR) and 185 bp ($2^{nd}$ PCR) fragments corresponding to a conserved region on the gag gene of CAEV. Positive ISH signals were detected in the brain and liver. In conclusion, significant histopathological findings included parenchymal infection in various organs, including the lung, liver, brain, joint, and mammary gland were noted in the CAEV infected dairy goat. ISH can help confirm the diagnosis of CAE in formalin-fixed samples.

• 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.

• Journal of Photoscience
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• v.9 no.3
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• pp.33-36
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• 2002

This study describes RT-PCR and in situ hybridisation protocols, and the immunohistochemical detection method that we have developed to detect and localise cells that express HO-1 in the skin. We found that HO-1 mRNA was absent in normal mouse skin, but after UVA irradiation HO-1 mRNA was expressed in the dermal fibroblasts, and strongly in basal epidermal cells. HO-1 protein was also induced strongly in dermal fibroblasts, and also in epidermal cells. In addition, the HO substrate heme was reduced in skin microsome at 72 hrs post UVA (when HO activity is high). At the same time, the HO products bilirubin and iron levels were elevated in the cutaneous tissue. Thus in addition to a dermal response, there appears to be an epidermal HO response to UVA in vivo that may be relevant for immune modulation by UVA radiation.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.41 no.1
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• pp.11-18
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• 2015

Objectives: The goal of this study was to determine the correlation of clinicopathological factors and the up-regulation of vascular endothelial growth factor (VEGF) expression in oral squamous cell carcinoma. Materials and Methods: Immunohistochemical staining of VEGF and quantitative real-time polymerase chain reaction (RT-PCR) of VEGF mRNA were performed in 20 specimens from 20 patients with oral squamous cell carcinoma and another 20 specimens from 20 patients with carcinoma in situ as a controlled group. Results: The results were as follows: 1) In immunohistochemical study of poorly differentiated and invasive oral squamous cell carcinoma, high-level staining of VEGF was observed. Significant correlation was observed between immunohistochemical VEGF expression and histologic differentiation, tumor size of specimens (Pearson correlation analysis, significance r>0.6, P<0.05). 2) In VEGF quantitative RT-PCR analysis, progressive cancer showed more VEGF expression than carcinoma in situ. Paired-samples analysis determined the difference of VEGF mRNA expression level between cancer tissue and carcinoma in situ tissue, between T1 and T2-4 (Student's t-test, P<0.05). Conclusion: These findings suggest that up-regulation of VEGF may play a role in the angiogenesis and progression of oral squamous cell carcinoma.

• Proceedings of the Korean Society of Veterinary Pathology Conference
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• 2003.10a
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• pp.26-26
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• 2003

Hepatitis E virus (HEV) has been recognized as a major cause of enterically transmitted non-A, non-B hepatitis in many developing countries. The taxonomy of HEV is not clear and the virus remains unclassified. The objective of this study was to optimize conditions and procedures to detect swine HEV in formalin-fixed, paraffin-embedded tissues by nested RT-PCR and compare this detection method with in situ hybridization. (omitted)

• Proceedings of the KSAR Conference
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• 2002.06a
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• pp.86-86
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• 2002

To examine the feasibility of using a sperm vector system for gene transfer, we have investigated the binding and the uptaking of foreign DNA into the sperm nucleus by PCR, in situ hybridization and LSC. We have also examined the transportation of exogenous DNA into oocytes by immunofluorescene via PCR. Sperm cells were incubated with DNA/liposome complexes (1:4 ratio) in fertilization medium with BSA or without BSA. (omitted)

• Journal of Aquaculture
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• v.18 no.3
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• pp.207-214
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• 2005

We report on the diagnosis, pathology, and taxonomy of Perkinsus sp. infection in Manila clams (Ruditapes philippinarum) from Korean waters. Amplimers were designed from internal portions of the non-transcribed spacer (NTS) of P. atlanticus for molecular diagnosis of Perkinsus infection. PCR-based identification methods and an in situ hybridization assay were developed for detection of Perkinsus sp. in live tissues as well as in histological preparations. Hybridization signals were observed around the nucleus of trophozoites. Positive results from PCR and in situ hybridization indicated that Korean Perkinsus sp. is genetically identical with P. atlanticus reported in Europe, which is currently synonymous with P. olseni reported from Australia. Microscopic morphological features of different lift stages of Perkinsus sp. appeared very similar to those of P. atlanticus. Severely infected clams often exhibited white nodules on their mantles and gills as a consequence of inflammation. In lightly to moderately infected clams, Perkinsus sp. was mainly found in gill tissues, whereas the protozoan parasites were found in digestive tracts, gonadal tissues, and foot tissues of heavily infected clams. It is likely that the gills are the portal of the infection and that P. olseni spreads to other tissues as the infection advances. In conclusion, by considering the taxonomic priority of P. olseni, Korean Perkinsus sp. is accepted as P. olseni. P. olseni appears to be common on tidal flats on the western and southern Korean coasts and is considered to be a pathogen capable of causing mass mortality of clams.

• Korean Journal of Animal Reproduction
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• v.20 no.3
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• pp.323-331
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• 1996

Predetermination of sex in mammalian species has many aspects of application including the prenatal diagnoses of genetic disorders in humans and sex-selected breeding programs in the animal industry. Embryos sexing can be carried out using the polymerase chain reaction (PCR) to amplify specific sequences present in the sex chromosomes, or by fluorescent in situ hybridization (FISH) of specific probes to the X and Y chromosomes. A 3.3 kb porcine male-specific DNA fragment (pEM39) was cloned previously in our laboratory. In this study, FISH and PCR methods were employed to examine if the pEM39 can be used a sex-specific DNA probes Porcine ovaries were obtained from a local slaughter house and oocytes collected. All oocytes were subjected to in vitro maturation followed by 1n vitro fertilization. Parthenogenetically activated embryos were served as a negative control. Embryonic samples were collected at the 2-cell stages and PCR was performed to analyze DNA. Among 10 embryos examined, four embryos were identified as males and six were females. The cloned male-specific DNA fragment showed male-specificity for the cells in the liver tissue and the porcine early embryos by FISH. It was also demonstrated that the cloned male-specific DNA is localized on the hetero chromatic region of the long arm in the Y chrom-osome (Yq) as shown by the FISH and karyotyping. The results suggest that the cloned male-specific DNA fragment may be useful for predetermination of sex with a few embryonic cells. The porcine male-specific sequence can be a reliable index for embryo sexing by PCR.