• 한국임상수의학회지
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• 제20권2호
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• pp.150-154
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• 2003

진주 지역에서 1000 여두의 생후 1주령 포유 자돈군에 심한 설사증이 발생하였던바 소장조직으로부터 RNA를 추출한 후 reverse transcription-polymerase chain reaction을 실시하였던 결과, porcine epidemic diarrhea (PED) virus (PEDV)의 N 유전자가 검출되어 PED로 진단되었다. Trypsin (10ug/ml)을 첨가한 배지에서 Vero 세포를 배양하면서 장조직으로부터 바이러스 분리배양을 시도하였던 결과 2회의 blind passage 후에 PEDV를 분리할 수 있었다. 따라서 이 분리주를 Chinju99로 명명하였으며, Chinju99주의 생물학적 및 물리화학적 성상을 조사하였던 결론은 다음과 같다. Chinju99주는 전자현미경 소견에서 비정형의 타원형 입자로서 표면에 spike 구조를 가지고 있었으며, Vero 세포내 배양시에 점진적으로 원형변성, 합포체형성 등의 세포변성 소견을 나타내었다. 또한 20% ether, 5% chloroform에서 불안정하였으며 pH 4-7의 산성조건에서는 안정성을 나타내었다. 동시에 $50^{\circ}C$에서 180분간 처리 후에도 감염성이 유지되었고 sucrose 용액에서의 부유밀도가 1.180g/ml으로서, coronavirus의 전형적인 생물학적 및 물리화학적 성상을 나타내었다.

• 한국임상수의학회지
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• 제31권4호
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• pp.319-321
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• 2014

12년령 접종 수컷 개가 전북대학교 수의과 대학에 왼쪽 편측성의 유루증으로 내원하였다. 확대경을 통한 안검사에서 왼쪽 안검 결막에 매우 작은 덩어리가 있는 것이 발견되었다. 이 조직을 외과적으로 제거하여 조직검사를 실시한 결과 편평상피 세포에 바이러스성 변화가 없는 중등도의 유두세포 증식이 관찰되었다. 조직 검사를 바탕으로 비바이러스성 편평세포 유두종으로 진단하였다. 조직 제거 후에 환자의 유루증은 완전히 해결되었다. 본 증례는 개에서 결막의 비바이러스성 편평세포 유두종에 의해 발생한 유루증을 치료한 것이다.

• 동국한의학연구소논문집
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• 제1권
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• pp.209-236
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• 1992

The microculture XTT antiviral assay method is used to quantitate HIV-1 induced cytopathic effects as modulated by test substances. This relatively simple assay facilitated the safe and rapid determination of in vitro antiviral activity of selected chemicals as well as direct cytotoxicity. This experiment also confirmed that this system measures infection and subsequent viral replication in target cells and XTT formazan formations correlated with the accumulation of extracellular virions, as measured by quantitative HIV-1 induced syncytium foramtion. The present results with Glycyrrhizin using this in vitro culture system demonstrated that effective dose, EC50(the concentration at which increases XTT formazan production in infected cultures to 50% of that in untreated, uninfected controls) was 250ml. As comparison, AZT was included in this experiment and demonstrated that EC50 AZT of was 0.05g/ml, approximately 5,000 times more potent than Glycyrrhizin based on EC50 ratio's alone. However, this potency is limited by severe cytotoxicity of AZT, while Glycyrrhizin is approximately 16 times less toxic(IC50 of Glycyrrhizin 800 and AZT 51 g/ml). While AZT's anti-HIV-1 viral activity is mediated by inhibition of reverse transcriptase of the virus, Glycyrrhizin faild to demonstrate any inhibitory activity against reverse transcriptase. Further study is necessary in order to understand the precise mechanisms of Glycyrrhizin action against HIV-1 viruses. Althouth Glycyrrhizin is less effective antiviral agent than AZT, much less toxicity of Glycyrrhizin is desirable in terms of chronic treatment. Combination treatment of AZT and Glycyrrhizin may be therapeutically beneficial. Clinical effectiveness of two drug combination therapy for AIDS patient is unknown at this time. However, this experimental investigation presents the scientific rational basis for such therapeutic approach.

• 한국어병학회지
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• 제30권2호
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• pp.71-78
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• 2017

The glycoprotein of novirhabdoviruses is known to play a critical role in the determination of host specificity. Viral hemorrhagic septicemia viruses (VHSVs) in different genotypes have different glycoprotein sequences and show different preferences for specific cell lines. In this study, to know whether the glycoprotein is solely responsible for the host cell preference of VHSV, a recombinant VHSV expressing vesicular stomatitis virus (VSV) glycoprotein instead of VHSV IVa glycoprotein (rVHSV-VSV-G) was generated by reverse genetics and inoculated into several fish cell lines, then, cytopathic effect (CPE) and viral growth caused by rVHSV-VSV-G infection were compared with those caused by rVHSV-wild that was previously generated and has the same genomic sequence with wild-type VHSV except a few nucleotides. The plaque numbers of rVHSV-VSV-G were significantly higher in EPC, BF-2 and GF cells than those of rVHSV-wild. However, in HINAE cells (originated from olive flounder), rVHSV-VSV-G titer was significantly lower than rVHSV-wild titer, and both recombinant VHSVs were not grown well in CHSE-214 cells. Although statistical significances were detected in the titers between rVHSV-wild and rVHSV-VSV-G in several cell lines, the cell line-preference order of rVHSV-VSV-G was not different from that of rVHSV-wild. These results suggest that the replacement of VHSV glycoprotein may not completely change host cell preference, and other regions of VHSV might also involve in the determination of host cell preference.

• 한국미생물·생명공학회지
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• 제32권2호
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• pp.179-183
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• 2004

2002년도 부산지역의 병ㆍ의원으로부터 의뢰된 바이러스성 뇌수막염으로 의심되는 환자의 가검물을 대상으로 바이러스 분리를 시도한 결과 703건의 검체 중 83건의 장내바이러스를 분리하였다. 이 중 echovirus 2, 3, 6, 7, 9, 13, 25, 29, 30 혈청형이 72건으로 가장 많았고, coxsackievirus B3과 B4혈청형이 11건으로 예전에 비해 보다 다양한 경향을 나타내었다. 월별발생 양상은 4월부터 11월까지 넓은 발생 분포를 나타내었지만 특히 6월과 7월에 가장 발생률이 높았다. Echovirus와 coksackievirus는 Veroft HEp-2 세포주에서 강한 병변 효과를 나타내었다. 연령별 및 성별 감염현황은 0-10세 사이의 어린이가 주로 발생하였고, 여아에 비해 남아의 비율이 비교적 높게 나타내었다. 전자현미경으로 촬영한 echovirus 및 coxsackievirus의 형태학적 양상은 모두 envelope가 없고 크기가 30∼35nm로 아주 작은 구형의 특징을 나타내었다. 세포병변 효과가 나타난 세포배양액에 대하여 nested PCR을 수행한 결과 echovirus 및 coxsackievirus 모두 437 bp위치에 단일 띠를 나타내었으며, serotype은 국립보건원 소화기바이러스과에 의뢰하여 확인 동정하였다.

• 한국어병학회지
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• 제22권3호
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• pp.229-237
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• 2009

The causative agent for the tunic softness syndrome of the cultured ascidian Halocynthia roretzi from Jan 1999 to Feb 2009 was identified using virus isolation and polymerase chain reaction (PCR). The pathogenicity of the isolated virus MABV UR-1 strain was determined by experimental infection trials. The cytopathic effects was observed in CHSE-214 cell line at a level 5.1% (4/78) in normal ascidian and 1.8% in abnormal ascidian showing tunic softness syndrome signs. MABV gene was detected in 16.8% (18/107) of normal and 13.1% (5/38) of abnormal organisms by PCR. The ratio of MABV isolation and gene detection was similar level in normal and soft tunic diseased ascidian. Based on the VP2/NS junction region sequences, eight strains of virus isolated from ascidian, were included in the same genogroup with MABV which is originally isolated in wide ranges of marine fish and shellfish species. The UR-1 strain caused 60% mortality (36.5% mortality in control group) by immersion infection and 37% mortality (same mortality in control group) in injection infection indicating no significant differences in infected and control groups. These results suggest that ascidian can act as reservoir of the MABV, and this virus is not directly related with the ascidian mortality.

• 한국가금학회지
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• 제40권3호
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• pp.243-252
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• 2013

Among the 16 hemagglutinin (HA) subtypes of avian influenza virus (AIV), only the H5 and H7 subtypes have caused highly pathogenic avian influenza (HPAI) in poultry. However, most H5 or H7 subtype viruses are categorized as low pathogenic avian influenza (LPAI). Some AIVs, including the H5 and H7 HPAI viruses, have shown the ability to infect humans directly. In this study, we describe the biological and molecular characterization of an H5N3 AIV (SBD/KR/KNU SYG06/06) isolated from spot-billed duck (Anas poecilorhyncha) in Korea. A phylogenetic analysis of the eight viral genes showed that the SBD/KR/KNU SYG06/06 isolate belongs to the Eurasian lineage and that the SBD/KR/KNU SYG06/06 isolate was clearly different from HPAI H5N1 strains, including human isolates and the Italian HPAI H5N2 strains. Additionally, no relationship was found between SBD/KR/KNU SYG06/06 and the Korean HPAI H5N1 isolates. The SBD/KR/ KNU SYG06/06 isolate had avian specific receptor binding site residues in the HA protein and the four C-terminal amino acids in the NS1 protein. The HA protein of the SBD/KR/KNU SYG06/06 isolate exhibited the typical LPAI motif at the cleavage site and this virus produced no cytopathic effects in MDCK cells without trypsin. Given these results, we suggest that the H5N3 AIV isolated from the spot-billed duck should be considered an LPAI virus and should have no pathogenic effect in humans.

• Fisheries and Aquatic Sciences
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• 제13권1호
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• pp.79-83
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• 2010

Since 1998, a new viral disease with high mortality has been consistently recorded in Korea in cultured carp, Cyprinus carpio. In this study, we investigated an epizootic of the disease that caused high mortality rates in carp obtained from 11 farms in Korea between 1999 and 2007. Assessment of koi herpesvirus (KHV) levels in diseased carp was carried out to determine if this virus was the etiologic agent of disease in this instance. High mortality rates in carp were recorded mainly in the spring and autumn at water temperatures between $19^{\circ}C$ and $24^{\circ}C$. Diseased fish typically showed surface discoloration, with a thick opaque mucus covering the body and gills. Protozoan parasites and bacteria were recovered from 7/29 (24%) and 2/26 (8%) of fish, respectively. Evidence of viral infection was marked; cytopathic effects (CPEs), characterized by cell rounding and an extended cytoplasm in fathead minnow (FHM) cells, were detected in 40/41 fish (98%). A high mortality rate (80%) resulted when supernatants of cell cultures showing CPEs were applied to previously healthy fish. KHV was detected by polymerase chain reaction in 6/41 fish (15%), but was not detected in supernatants obtained from cell cultures showing CPEs. These results suggest that KHV may not be the etiologic agent of the high mortality occurring among cultured carp in Korea; therefore, some other-as yet unidentified-infective agent must be responsible.

• Molecules and Cells
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• 제44권9호
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• pp.688-695
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• 2021

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic has become a global health concern. Various SARS-CoV-2 vaccines have been developed and are being used for vaccination worldwide. However, no therapeutic agents against coronavirus disease 2019 (COVID-19) have been developed so far; therefore, new therapeutic agents are urgently needed. In the present study, we evaluated several hepatitis C virus direct-acting antivirals as potential candidates for drug repurposing against COVID-19. Theses include asunaprevir (a protease inhibitor), daclatasvir (an NS5A inhibitor), and sofosbuvir (an RNA polymerase inhibitor). We found that asunaprevir, but not sofosbuvir and daclatasvir, markedly inhibited SARS-CoV-2-induced cytopathic effects in Vero E6 cells. Both RNA and protein levels of SARS-CoV-2 were significantly decreased by treatment with asunaprevir. Moreover, asunaprevir profoundly decreased virion release from SARS-CoV-2-infected cells. A pseudoparticle entry assay revealed that asunaprevir blocked SARS-CoV-2 infection at the binding step of the viral life cycle. Furthermore, asunaprevir inhibited SARS-CoV-2 propagation in human lung Calu-3 cells. Collectively, we found that asunaprevir displays broad-spectrum antiviral activity and therefore might be worth developing as a new drug repurposing candidate for COVID-19.

• 대한수의학회지
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• 제61권2호
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• pp.13.1-13.8
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• 2021

Mammalian reovirus (MRV) causes respiratory and intestinal disease in mammals. Although MRV isolates have been reported to circulate in several animals, there are no reports on Korean MRV isolates from wildlife. We investigated the biological and molecular characteristics of Korean MRV isolates based on the nucleotide sequence of the segment 1 gene. In total, 144 swabs from wild animals were prepared for virus isolation. Based on virus isolation with specific cytopathic effects, indirect fluorescence assays, electron microscopy, and reverse transcription-polymerase chain reaction, only one isolate was confirmed to be MRV from a Korean roe deer (Capreolus pygargus). The isolate exhibited a hemagglutination activity level of 16 units with pig erythrocytes and had a maximum viral titer of 10^5.7 50% tissue culture infectious dose (TCID₅₀)/mL in Vero cells at 5 days after inoculation. The nucleotide and amino-acid sequences of the partial segment S1 of the MReo2045 isolate were determined and compared with those of other MRV strains. The MReo2045 isolate had nucleotide sequences similar to MRV-3 and was most similar (96.1%) to the T3/Bat/Germany/342/08 strain, which was isolated in Germany in 2008. The MReo2045 isolate will be useful as an antigen for sero-epidemiological studies and developing diagnostic tools.