• 한국수산과학회지
• /
• 제51권3호
• /
• pp.328-331
• /
• 2018

We developed and subsequently characterized mouse monoclonal antibodies (MAbs) against nervous necrosis virus (NNV, RGNNV genotype). We established six hybridoma clones secreting MAbs against NNV antigen: 2B1, 2B11, 2C12, 13C1-1, 13C1-2 and 14D11. All six MAbs belonged to the IgG2a isotype with a kappa light chain and their reactivity recognized against the 41 kDa coat protein of NNV by Western blot analysis. The affinity constants of the six MAbs were measured by enzyme-linked immunosorbent assay (ELISA). All six MAbs reacted with two NNV isolates (SgNag05 and Gemunodo06), while no reactivity was observed with five know fish viruses, namely marine birnavirus, infectious pancreatic necrosis virus, viral hemorrhagic septicemia virus, hirame rhabdovirus, and infectious hematopoietic necrosis virus. Moreover, high ELISA optical density (OD) values (0.87-1.42) were observed in the brain tissues of NNV-infected sevenband grouper, while low OD values (less than 0.12) were recorded in the brain tissues of uninfected fish. These results suggest that these six MAbs are highly competent and useful for the detection of NNV with the RGNNV genotype.

• 한국동물위생학회지
• /
• 제20권4호
• /
• pp.331-338
• /
• 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.

• Journal of Microbiology and Biotechnology
• /
• 제24권7호
• /
• pp.979-986
• /
• 2014

Plant pathogenic RNA viruses are present in a variety of plant-based foods. When ingested by humans, these viruses can survive the passage through the digestive tract, and are frequently detected in human feces. Kimchi is a traditional fermented Korean food made from cabbage or vegetables, with a variety of other plant-based ingredients, including ground red pepper and garlic paste. We analyzed microbial metatranscriptome data from kimchi at five fermentation stages to identify plant RNA virus-derived sequences. We successfully identified a substantial amount of plant RNA virus sequences, especially during the early stages of fermentation: 23.47% and 16.45% of total clean reads on days 7 and 13, respectively. The most abundant plant RNA virus sequences were from pepper mild mottle virus, a major pathogen of red peppers; this constituted 95% of the total RNA virus sequences identified throughout the fermentation period. We observed distinct sequencing read-depth distributions for plant RNA virus genomes, possibly implying intrinsic and/or technical biases during the metatranscriptome generation procedure. We also identified RNA virus sequences in publicly available microbial metatranscriptome data sets. We propose that metatranscriptome data may serve as a valuable resource for RNA virus detection, and a systematic screening of the ingredients may help prevent the use of virus-infected low-quality materials for food production.

• 한국동물위생학회지
• /
• 제33권2호
• /
• pp.105-111
• /
• 2010

Rabies virus which belongs to the genus Lyssavirus of the family Rhabdoviridae is known as a highly neurotropic virus and causes fatal encephalitis accompanied by severe neurological symptoms in almost all mammals, including humans. In this study, monoclonal antibodies (MAbs) against rabies virus were produced, characterized and applications of MAbs as diagnostic reagents were assessed Spleen and inguinal lymph node cells from Balb/c mouse immunized with purified rabies virus were fused with SP2/O myeloma cells using polyethylene glycol (PEG) and hybridoma cells producing rabies virus-specific MAbs were screened by an indirect fluorescent antibody test. A total of ten MAbs were produced against rabies virus. The protein specificity and neutralizing activity of MAbs were determined by Western blot analysis and fluorescent antibody virus neutralization test, respectively. As a result, two MAbs, 5G3 and 6H4 had specificity for nucleoprotein (N protein) and two other MAbs, 5B1 and 5C1 had neutralizing activity for rabies virus. Some MAbs recognized the rabies virus-infected bovine brain stem cells by immunohistochemistry (IHC) assay. In conclusion, it was confirmed that MAbs produced in this study were rabies virusspecific and could be used as reliable diagnostic reagents for the detection of rabies virus.

• Perspectives in Nursing Science
• /
• 제7권1호
• /
• pp.50-54
• /
• 2010

The studies of M. Colombo (1989) and W. Lange (1992) showed that 30~40% of people became chronic after suffering from hepatitis B virus (HBV) and C virus (HCV) infection, and about 50% of the chronic cases transformed into primary liver cancer. There have been few studies done in Mongolia on hepatitis infection among health professionals, particularly in nurses. In a study done by Chimedsuren (8), the study showed that 19.4% of people with identified surface hepatitis B antigen (HBsAg) and antibodies to hepatitis C virus and 8% of people with the identified nucleotide of RNA for the hepatitis C virus (polymerase chain reaction) had an acute form of hepatitis C. Studies on the hepatitis virus genome damaging effect on liver cells showed that genotype 8 (A, B, C, D, E, F, G, TTV) had the most damaging effect on liver cells (Hahn and Faeka, 2007). Several studies have shown a relationship between hepatitis B virus infection and a lack of compliance regarding safety regulations and rules by medical personnel. Results of a study from the Maternal and Child Health Research Center showed that tests done to detect hepatitis B virus antigen and antibodies to C virus did not reveal anything. Both antigen and antibodies in 69% cases did not show, and separately, B virus and antibodies to hepatitis C virus were identified in 13% and 9%, respectively. Results of the tests taken from health personnel in Shastin Central Hospital showed that in 76% of the cases, the B virus antigen with C virus antibodies was not identified. In 8% of the cases, the B virus antigen was present on its own. The combination of B the virus antigen and C virus antibodies were present in 8% of nurses and doctors, respectively. 82% of the cases had negative results for the detection of a combination of B virus antigen and C virus antibodies taken from health personnel from the State Central Clinical Hospital whereas the B virus antigen and C virus antibodies by themselves were present in 7% and 14% of the cases, respectively. Combined cases of the B virus antigen and C virus antibodies were identified in 4% of the personnel. Results of the tests taken from the health personnel in the Hospital of the Ministry of Justice and Internal Affairs showed that in 79% of the cases, the B virus antigen with C virus antibodies were not identified. Separately, the B virus and antibodies to hepatitis C virus were identified in 8% and 13% of the cases, respectively.

• 한국생물공학회:학술대회논문집
• /
• 한국생물공학회 2003년도 생물공학의 동향(XIII)
• /
• pp.99-99
• /
• 2003

• Molecular & Cellular Toxicology
• /
• 제3권sup4호
• /
• pp.37.2-37.2
• /
• 2007

• BMB Reports
• /
• 제38권6호
• /
• pp.683-689
• /
• 2005

Antibody to hepatitis B surface antigen (HBsAb) is the important serological marker of the hepatitis B virus (HBV) infection. Conventionally, the hepatitis B surface antigen (HBsAg) obtained from the plasma of HBV carriers is used as the diagnostic antigen for detection of HBsAb. This blood-origin antigen has some disadvantages involved in high cost, over-elaborate preparation, risk of infection, et al. In an attempt to explore the suitable recombinant HBsAg for the diagnostic purpose, the HBV S gene was expressed in Pichia pastoris and the product was applied for detection of HBsAb. Hepatitis B virus S gene was inserted into the yeast vector and the expressed product was analyzed by sodium dodecyl sulphate polyacrolamide gel electrophoresis (SDS-PAGE), immunoblot, electronic microscope and enzyme linked immunosorbent assay (ELISA). The preparations of synthesized S protein were applied to detect HBsAb by sandwich ELISA. The S gene encoding the 226 amino acid of HBsAg carrying ahexa-histidine tag at C terminus was successfully expressed in Pichia pastoris. The His-Tagged S protein in this strain was expressed at a level of about 14.5% of total cell protein. Immunoblot showed the recombinant HBsAg recognized by monoclonal HBsAb and there was no cross reaction between all proteins from the host and normal sera. HBsAb detection indicated that the sensitivity reached 10 mIu (micro international unit)/ml and the specificity was 100% with HBsAb standard of National Center for Clinical Laboratories. A total of 293 random sera were assayed using recombinant S protein and a commercial HBsAb ELISA kit (produced by blood-origin HBsAg), 35 HBsAb positive sera and 258 HBsAb negative sera were examined. The same results were obtained with two different reagents and there was no significant difference in the value of S/CO between the two reagents. The recombinant HBV S protein with good immunoreactivity and specificity was successfully expressed in Pichia pastoris. The reagent for HBsAb detection prepared by Pichia pastoris-derived S protein showed high sensitivity and specificity for detection of HBsAb standard. And a good correlation was obtained between the reagent produced by recombinant S protein and commercial kit produced by blood-origin HBsAg in random samples.

• The Plant Pathology Journal
• /
• 제29권1호
• /
• pp.99-104
• /
• 2013

To detect five plant viruses (Beet black scorch virus, Beet necrotic yellow vein virus, Eggplant mottled dwarf virus, Pelargonium zonate spot virus, and Rice yellow mottle virus) for quarantine purposes, we designed 15 RT-PCR primer sets. Primer design was based on the nucleotide sequence of the coat protein gene, which is highly conserved within species. All but one primer set successfully amplified the targets, and gradient PCRs indicated that the optimal temperature for the 14 useful primer sets was $51.9^{\circ}C$. Some primer sets worked well regardless of annealing temperature while others required a very specific annealing temperature. A primer specificity test using plant total RNAs and cDNAs of other plant virus-infected samples demonstrated that the designed primer sets were highly specific and generated reproducible results. The newly developed RT-PCR primer sets would be useful for quarantine inspections aimed at preventing the entry of exotic plant viruses into Korea.

• 한국동물위생학회지
• /
• 제38권2호
• /
• pp.107-116
• /
• 2015

In this study, we developed a rapid, sensitive and specific reverse-transcriptase loop-mediated isothermal amplification (RT-LAMP) assay for detection of swine influenza viruse (SIV) including major subtypes of swine influenza viruses H1N1, H1N2 and H3N2, and a novel subtype of influenza A virus that accidentally infected in pig population. The RT-LAMP was completed in 40 min at $58^{\circ}C$ and the sensitivity of the RT-LAMP ($1copy/{\mu}L$) was 10-fold higher than conventional reverse transcription-polymerase chain reaction (RT-PCR) ($10copy/{\mu}L$) and the same to real time RT-PCR ($1copy/{\mu}L$). Also, the result of the RT-LAMP can be confirmed without any detection system. Therefore, the RT-LAMP could be a alternative diagnostic method for SIV detection in national SIV monitoring system and clinical diagnostic laboratory in the future.