• 한국수산과학회지
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• 제51권3호
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• pp.328-331
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• 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.

• 한국식품위생안전성학회지
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• 제9권4호
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• pp.221-228
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• 1994

The quantitative detection of ochratoxin A (OT-A) in the traditional fermented foods were investigated to develop the analytical procedures, Enzyme-Linked Immunosorbent Assay(ELISA) and Chemiluminescence Immunoassay(CIA). Products used were divided into two groups: the first was the home-made 13 Doenjang, 12 Kanjang, and 14 Gochujang; and the second the traditional commercial products, 17 Deonjang and 11 Kanjang, which collected throughout the country. The standard curve for the quantitative determination of OT-A showed that the sensitivities in ELISA and CIA were upto the level of 20 pg/assay, and that the OT-A recovery rates were appeared to be more than 90%. The residual OT-A in the home-made products were 7.1$\pm$3.7 ng/g for Deonjang, 2.1$\pm$4.1 ng/g Kanjang were found in the traditional commercial products. Residual OT-A in the home-made products was comparatively far less than that of the traditional commercial products. At heat stability test of OT-A in the traditional fermented foods was found to be stable even at 121$^{\circ}C$ for 120 min.

• 대한의생명과학회지
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• 제10권3호
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• pp.219-230
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• 2004

This experiment was performed to examine the in vitro and in vivo affects of the two different haplotype strains of mice, BALB/c and C3H/HeN infected with Echinostoma hortense, and the manifestation of the profiles of cytokine in the splenocytes. In the in vitro experiment, the two mice's splenocytes were divided and stimulated with antigen of crude extracts and the antigen of excretory and secretory products of an adult warm and the manifestation of cytokine mRNA was verified with RT-PCR. As a result, the two different strains of mice both strongly manifested the Th2 cytokine rather than the Thl cytokine and in the case of the Th2 cytokine, the BALB/c mice manifested more strongly than the C3H/HeN mice. In the experiment using the ELISA method, the protem cytokine manifestation had the same result as the mRNA experiment. In the in vivo experiment, the mice was infected via oral route with the metacercaria of the Echinostoma hortense and the manifestation of cytokine was verified by RT-PCR and ELISA and the results were the same as the in vitro experiment. Therefore, in the two strains of BALB/c and C3H/HeN, the C3H/HeN showed a higher susceptivity to the Echinostoma hortense.

• 한국동물위생학회지
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• 제44권1호
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• pp.27-33
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• 2021

Q fever is a worldwide zoonotic disease caused by Coxiella burnetii. Domestic ruminants are considered to be major source of human infection. The aim of this survey was to investigate seroprevalence of C. burnetii in ruminants in Gwangju area. A total of 1,000 samples (serum and lactoserum) were collected from 987 Korean native cattle, 5 Korean native goats, 2 beef cattle, 6 bulk-tank milk from each dairy farm in Gwangju area from January to October 2020 and analyzed by ELISA. The seroprevalence of C. burnetii in bulk-tank milk from each dairy farms was 50.0%. Korean black goat and beef cattle had negative antibody test results for C. burnetii. The seroprevalence of C. burnetii in Korean native cattle in Gwangju area was 7.1% and was higher in female (7.8%) than in male (3.4%) (P=0.024). The seroprevalence of C. burnetii in Korean native cattle appeared to increase with age (3.8% in 1 year-old, 7.1% in 3 year-old, and 10.7% in more than 5 year-old) (P<0.001). The seroprevalence of C. burnetii of Korean native cattle increased in spring and May was the highest in particular (P<0.001). As the distribution and density of tick-habitat are expected to increase due to climate crisis, this survey highlights the need for monitoring C. burnetii in domestic ruminants, including surveillance of C. burnetii infection in people working for livestock industry.

• Journal of Pharmaceutical Investigation
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• 제31권3호
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• pp.191-196
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• 2001

Biodegradable polymeric microspheres were studied for their usefulness as carriers for the delivery of vaccine antigens. However, protein antigen could be denatured during microencapsulation processes due to the exposure to the organic phase and stress condition of cavitation and shear force. Therefore this study was carried out to re-evaluate the degree of protein denaturation during microencapsulation with poly(lactide-co-glycolide) (PLGA) copolymer. PLGA microspheres containing ovalbumin (OVA), prepared by W/O/W multiple emulsification method, were suspended in pH 7.4 PBS and incubated with shaking at $37.5^{\circ}C$. Drug released medium was collected periodically and analyzed for protein contents by micro-BCA protein assay. In order to evaluate the protein integrity, release medium was subjected to the analyses of SDS-PAGE and size exclusion chromatography (SEC). And enzyme-linked immunosorbent assay (ELISA) was introduced to measure the immunoreactivity of entrapped OVA and to get an insight into the three-dimensional structure of epitope. The structures of entrapped protein were not affected significantly by the results of SDS-PAGE and SEC. However, immunoreactivity of released antigen was varied, revealing the possibility of protein denaturation in some microspheres when it was evaluate by ELISA method. Therefore, in order to express the degree of protein denaturation, antigenicity ratio (AR) was obtained as follows: amount of immunoreactivity of OVA/total amount of OVA released ${\times}100(%)$. ELISA method was an efficient tool to detect a protein denaturation during microencapsulation and the comparison of AR values resulted in more accurate evaluation for immunoreactivity of entrapped protein.

• 한국미생물·생명공학회지
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• 제16권2호
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• pp.168-173
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• 1988

곤충치사 독소를 생성하는 Bacillus thuringiensis serovar. kurstaki HD1과 HD73 균주에 대한 내독소 단백질의 특성을 규명하기 위하여 독소단백질의 항체 생산성, 항원 안정성, 정량 및 교차반응실험을 수행하였다. 곤충치사 내독소 단백질에 대한 항체는 토끼 마리당 약 2mg의 내독소 항원을 4회 주사했을 때 급격히 증가하여 6~7회 투여시에 최대치의 항체가 생성되었다. 내독소의 항원 안정성을 ELISA로 조사했을 때 5일 이내는 안정하였으나 9일 이후는 급격히 감소하였다. 간접 ELISA에 의한 B. t. k. HD1과 HD73 내독소의 정량 감도는 각각 50$\mu\textrm{g}$/$m\ell$, 400ng/$m\ell$이었다. HD1과 HD73 균주의 내독소 단백질에 대한 항혈청은 B. t. serovar israelensis 와 침강반응이 일어나지 않았다. B. t. k. 균주의 동일 아종간 내독소 단백질에 대한 항체의 교차반응에서 HD1 내독소 항체는 HD73 균주의 내독소 항원과 부분적으로 반응하였으나 HD73 내독소 항체는 HD1 항원과 100% 반응하였다.

• Perinatology
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• 제29권4호
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• pp.165-169
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• 2018

Objective: Highly sensitive haptoglobin measurement should be used in neonates because the haptoglobin concentration in neonates is lower than that of adults. The aim of this study was to establish the reference values of haptoglobin levels in the cord blood of uninfected neonates. Methods: The cord blood of 29 preterm and 51 term babies was collected, and data from the mother and the newborn were recorded. The haptoglobin concentrations of 80 cord blood samples were simultaneously measured by enzyme-linked immunosorbent assay (ELISA; Assaypro, St Charles, MO, USA) and immunoturbidimetry assay (Roche Diagnostics, Basel, Switzerland). C-reactive protein (CRP) was also measured by immunoturbidimetry assay (Roche Diagnostics, Switzerland). Results: Mean values of CRP and ELISA haptoglobin were not significantly different between preterm and term babies. The 2.5 percentile and 97.5 percentile values of ELISA haptoglobin concentration were as follows: 80 neonates, 0.01 mg/dL and 0.59 mg/dL; 29 preterm babies, 0.08 mg/dL and 0.18 mg/dL; and 51 term babies, 0.07 mg/dL and 0.23 mg/dL. There were no differences in ELISA haptoglobin concentration according to maternal underlying diseases, delivery method, usage of antibiotics or steroids before delivery, gestational age, gender of baby, or twin gestation. Conclusion: A highly sensitive haptoglobin method should be used to determine the haptoglobin concentration in Korean newborns because the reference values of cord blood haptoglobin concentration in Korean newborns are less than the lower detection limit for commonly used immunoturbidimetric haptoglobin measurement methods.

• The Plant Pathology Journal
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• 제19권5호
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• pp.260-265
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• 2003

Gummy stem blight caused by Didymella bryoniae occurs exclusively in cucurbits. This fungus has been known not to produce its pycnidium in vitro unless irradiated. In this study, cultural conditions for the mass-production of pycnidiospore by Metal Halide (MH) lamp irradiation were maximized. The mycelia were cultured at $26^{\circ}C$ on PDA for 2 days under dark condition, and then the plate was illuminated with MH lamp continuously for 3-4 days at $26^{\circ}C$. Results show that a great number of pycnidia were simultaneously formed. The pycnidiospores produced were then used as immunogen. Fusions of myeloma cell (v-653) with splenocytes from immunized mice were carried out. Two hybridoma cell lines that recognized the immunogen D. bryoniae were obtained. One monoclonal antibody (MAb), Dbl, recognized the supernatant while another MAb, Db15, recognized the spore. Two clones were selected which were used to produce ascite fluid of the two MAb, Dbl and Db15, the immunotypes of which were identified as IgG1 and IgG2b, respectively. Titers of MAb Dbl and MAb Db15 were measured and the absorbance exceeded 0.5 even at a $10^{-5}$ dilution. The MAbs reacted positively with D. bryoniae but none reacted with other viral isolates, Cucumber mosaic virus and Cucumber green mottle mosaic virus. Sensitivity of MAb was precise enough to detect spore concentration as low as $10^{-3}$/well by indirect ELISA. Characterization of the MAbs Dbl, Db15 antigen by heat and protease treatments, which suggests that the epitope recognized by these two MAbs was glycoprotein.

• Asian-Australasian Journal of Animal Sciences
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• 제28권5호
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• pp.691-696
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• 2015

The objective of this study was to evaluate the quality of five commercial enzyme linked immunosorbent assay (ELISA) kits (A, B, C, D, and E) from different suppliers for detecting aflatoxin $B_1$ ($AFB_1$). $AFB_1$-free corn samples supplemented with different levels of $AFB_1$ (5, 10, and $20{\mu}g/kg$) were used as positive controls and 6 replicates of each control sample were tested to evaluate the accuracy and precision of these kits. In addition, we also evaluated the performance of these ELISA kits for $AFB_1$ in 30 feed samples, including corn, distillers dried grains with soluble, wheat samples, soybean meal, and poultry feed, which were verified by high performance liquid chromatography. Results showed that the coefficients of variation ranged from 1.18% to 16.22% in intra-plate and 2.85% to 18.04% in inter-plate for the determination of $AFB_1$. The half maximal inhibitory concentration for five kits ranged from 3.72 to $7.22{\mu}g/kg$. The quantitation limits of $AFB_1$ were all under the legal limit in China but somewhat inconsistent with kit instructions. Although the recovery rate of four of the five kits were either less than 90% or more than 110%, all these values were acceptable in practice. Two kits had high false positive rates (C and E). In conclusion, our results revealed that the qualities of five tested ELISA kits were significantly different.

• 한국식품과학회지
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• 제30권6호
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• pp.1409-1414
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• 1998

Zearalenone에 특이한 단크론성 항체를 개발하기위하여 형질세포종세포인 myeloma cell $(P3{\times}63Ag.V653)$과 zearalenone-oxime-bovine serum albumin (BSA) 결합체를 항원으로 면역시킨 BALB/c female mice의 비장세포를 융합시켜 hybridoma cell을 생산하였다. 그들의 항체가를 indirect competitive ELISA법으로 측정한 결과 zearalenone에 대하여 높은 친화력을 갖는 5세포주를 얻었다. 그중 Z-2-M26 hybridoma cell line이 생산하는 단크론성항체는 특히 zearalenone과 민감하게 반응하였고, ${\alpha}-zearalenol$과도 약간의 교차반응을 보였으나 ${\beta}-zearalenol,\;{\alpha}-zearalenol,\;{\beta}-zearalenol$ 및 DON과는 거의 반응하지 않았다. 따라서 본 실험에서 개발된 항체는 앞으로 zearalenone에 대한 민감하고 정량적인 효소면역측정법의 개발을 위한 중요한 면역시약으로 사용될 것으로 생각한다.