• Journal of Food Hygiene and Safety
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• v.18 no.2
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• pp.61-66
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• 2003

This study was conducted to produce the egg yolk Imunoglobulin (IgY) on Vibrio parahaemolyticus from immunized hen with lipopolysaccharide (LPS). Vibrio parahaemolyticus is considered as a potentially pathogenic bacteria, the causative agents of the gastroenteritis. According as the LPS antigens were injected into laying hens in order to produce antibodies against Vibrio parahaemolyticus in egg yolk. After chickens were immunized four times in 2 weeks interval and three times booster in 2 weeks interval, the profile of antibody Production was examined by ELISA. The Production of antibody in egg yolk was started in 1 week after the first immunization, reached peak in 7 weeks and maintained until 13 weeks later. The antibody titre in serum showed similar tendency as IgY. No significant difference in antibody titre when the titre compared to water diluted IgY and commercial IgY kit. Purified IgY reacted with only Vibrio parahaemolyticus, but other Vibrio species and food-borne pathogenic bacteria. In conclusion, we showed that it is possible to obtain a high antibody titre in chicken with quite low amounts of LPS antigen. These results suggested that egg yolk antibodies could be a good source for production of specific antibodies to pathogenic bacteria inducing epidemic gastroenteritis.

• Korean Journal of Clinical Laboratory Science
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• v.49 no.4
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• pp.395-400
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• 2017

Atopic dermatitis is a chronic inflammatory skin disease caused by a variety of genetic and environmental factors, dysregulation of immunological response, as well as dysfunction of the skin barrier proteins. The purpose of this study is to develop an ELISA kit suitable for evaluating the expression of skin barrier proteins. Proteins were obtained from the skin via AriNo and D-Squame patches. The efficiency of protein collection from the skin, using the Arino patch, was shown to be more effective than using D-Squame; while the efficiency of lysis using 0.1% Triton-X100 was higher than that of other lysis solutions, including 0.1 M Tris-HCL, 0.1% Tween-20, and 5 mM KOH. Recombinant skin barrier proteins, such as filaggrin and involucrin, were produced by molecular biological methods. Monoclonal antibodies against filaggrin and involucrin were produced by immunization of mice, fusion of spleen cells and myeloma cells, as well as a selection of antibody-producing hybridoma cells. The filaggrin expression in the skin of subjects suffering from atopic dermatitis was lower than that in normal mice. Involucrin expression was not altered between normal individuals and subjects with atopic dermatitis. These findings contribute to an elucidation of the importance of the skin barrier protein expression in atopic dermatitis and the development of a diagnostic kit for atopic dermatitis.

• Parasites, Hosts and Diseases
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• v.56 no.5
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• pp.437-446
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• 2018

To investigate the prevalence of Toxoplasma gondii in pork on the market in Korea, an in-house enzyme-linked immunosorbent assay for tissue fluid (CAU-tf-ELISA) was developed using a soluble extract of T. gondii RH strain tachyzoites. As the standard positive controls, the piglets were experimentally infected with T. gondii: Group A (1,000 cysts-containing bradyzoites), Group B (500 cysts-containing bradyzoites) and Group C ($1.0{\times}10^3$ or $1.0{\times}10^4$ tachyzoites). The CAU-tf-ELISA demonstrated infection intensity-dependent positivity toward tissue fluids with average cut-off value 0.15: 100% for Group A, 93.8% for Group B and 40.6% for Group C. When tissue-specific cut-off values 0.066-0.199 were applied, CAU-tf-ELISA showed 96.7% sensitivity, 100% specificity, 100% positive and 90.0% negative predictive values. When compared with the same tissue fluids, performance of CAU-tf-ELISA was better than that of a commercial ELISA kit. Of the 583 Korea domestic pork samples tested, anti-T. gondii antibodies were detected from 9.1% of whole samples and 37.9% from skirt meat highest among pork parts. In the 386 imported frozen pork samples, 1.8% (skirt meat and shoulder blade) were positive for anti-T. gondii antibodies. In Korea, prevalence of anti-T. gondii antibodies in the pork on retail markets appeared high, suggesting that regulations on pig farming and facilities are necessary to supply safe pork on the tables.

• Bulletin of the Korean Chemical Society
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• v.28 no.6
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• pp.929-935
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• 2007

The objective of this study was to develop a rapid, simple, and qualitative acetylcholinesterase (AChE)- detection kit, based on a modification of the Ellman and ELISA methods, for the detection of organophosphorus (OP) and carbamate (CB) pesticide. The developed kits were used to screen a large number of agricultural samples (spiked and real) for OP and CB pesticide residues. AChE was extracted from the heads of honeybees (Apis mellifera L.) using Triton X-100, and was purified through 3 steps: diethylaminoethylcellulose chromatography (DEAE), affinity chromatography and membrane filtering, and Mono-Q column chromatography. Epoxy-activated Sepharose 6B affinity chromatography was used for large-scale purification. The presence of OP and CB pesticide residues in agricultural samples was assayed on the basis of AchE inhibition value. The presence (6 bands) or absence of some colored bands on the test line indicated a negative or positive result, respectively. The limits of detection for measured organophosphorus (OP) and carbamates (CB) pesticide residues in standard pesticide solutions and fortified samples were ranged from 0.50 to 2.50 ppm and 0.50 to 4.75 ppm, respectively.

• BMB Reports
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• v.29 no.3
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• pp.192-199
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• 1996

A rapid and sensitive method has been developed to detect a-fetoprotein (AFP) in human serum by a two-site sandwich enzyme-linked immunosorbent assay (ELISA) with monoclonal antibodies (MAbs) for human AFP within 1 h. To obtain the most sensitive and reliable MAbs. 12 kinds of MAbs (HPJ1 to HPJ12) as a capture antibody and 4 kinds of horseradish peroxidase (HRP) conjugated MAbs as a tracer antibody were investigated. Among these, only HPJ 10-HRP conjugated HPJ 1 (HPJ 10-HPJ $1^*$) and HPJ 11-HRP conjugated HPJ 10 (HPJ 11-HPJ $10^*$) were chosen as candidates based on the linearity of the standard curve and the sensitivity of the assay. To further characterize these two pairs. MAbs against human AFP were purified from hybridoma cells. conjugated with HRP. and then characterized to optimize the two-site sandwich ELISA The HPJ 10-HPJ $1^*$ pair showed a sensitivity of 1 ng/ml and a better reproducibility than the HPJ 11-HPJ $10^*$ pair when the human sera were incubated at $37^{\circ}C$ for 30 min. The results obtained for 480 randomly selected human sera showed 0~20 ng/ml of AFP values for the normal human sera. To test the utility of our kit, AFP concentrations were determined for 951 human sera (including 85 normal sera, 480 random blood sera, 213 HBsAg-positives. 50 anti-HCV antibody positives. and 47 malignant diseases) and compared with other commercially available AFP detecting kits. These results show that the present two-site sandwich ELISA method is a rapid, sensitive, and reliable procedure for detecting AFP in human serum.

• Korean Journal of Clinical Laboratory Science
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• v.46 no.1
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• pp.12-16
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• 2014

Tetanus has high lethality and can cause serious complication, so it is very important to have a quick and exact checkup and treatment. In this study, we conducted a study about clinical characteristics and types of tetanus patients in Daegu Emergency Medical Center and we studied possibility of Tetanos Quick Stick (TQS) as a selective check up to treat tentanus by comparing the results of local checkup kit measured from tetanus quick stick developed to check up tetanus antibody titer and results measured from ELISA (Enzyme-Linked Immunosorbent Assay). The result of the study showed that tetanus happens more to males, and by looking at season, tetanus happened most in summer (from June-August) as 19 cases (45.3%), and when patients come to the emergency medical center, the diagnosis name was electrolyte imbalance 14 cases (33.3%), peripheral nerve 11 cases (26.2%), Meningitis 8 cases (19.0%), drug addiction 7 cases (16.7%), and the patients who are diagnosed as tetanus at the beginning of hospitalization was 2 cases (4.8%). The result of TQS usefulness by comparing with ELISA, in TQS, 42 people was positive and 478 people was negative. it was positive when the result was over 0.1 IU/mL, 48 people was ELISA positive while 472 people was negative. TQS checkup has accuracy of 98%, sensitivity of 83.3%, specificity of 99.5%, positive predictability of 95.2% and negative predictability of 98.3%. The evaluation of current immunity statuses of tetanus patients is available for TQS checkup, and it has an advantage of preventing side effects coming from the injection of unnecessary vaccine and immunoglobulin, and it is thought that it can give help to emergency checkup and treatment at the beginning.

• Korean Journal of Microbiology
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• v.39 no.3
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• pp.161-165
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• 2003

Ornithobacterium rhinotracheale (OR) is a recently described gram-negative rod-shaped bacterium associated with respiratory tract infection in poultry. In order to investigate current occurrence of OR infection and to evaluate antibiotic susceptibility, the prevalence of OR antibody in domestic chickens were examined and the minimal inhibitory concentrations(MICs) of 8 antibiotics for 11 OR isolates was determined. All isolates tested were mostly susceptible to three antibiotics, ampicillin (MICs ranging from 0.38 ${\mu}g$/ml to 2 ${\mu}g$/ml), tetracycline (MICs 0.094～3 ${\mu}g$/ml) and doxycycline (MICs 0.047～4 ${\mu}g$/ml) but resistant to genatmicin. Ciprofloxacin, norfloxacin, enrofloxacin, and ofloxacin gave most isolates inhibition only in case of a higher concentration (MICs ranged in most cases from 3 ${\mu}g$/ml to 48 ${\mu}g$/ml). Out of 188 chicken flocks including broilers, broiler breeders, and layers, seropositive flock to OR were detected in 5 broilers (4%), 17 broiler breeders (50%), and 16 layers (55.2%), using commercial OR enzyme-linked immunosorbent assay(ELISA) kits. It suggested that OR infection was widespreaded in poultry farms in Korea.

• Biomedical Science Letters
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• v.5 no.1
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• pp.1-10
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• 1999

This study was attempted to generate a monoclonal antibody against human $\alpha$-fetoprotein (AFP) and to produce an immunoassay, recognizing AFP in plasma and amniotic fluid. AFP was purified from human amniotic fluid and used to immunize mice. Spleens were taken from the mice and the cells were fused with mouse myeloma cells (Sp2/0-Ag-14) for the production of monoclonal antibodies by employing the hybridoma technology. As a result, a hybridoma cell line producing anti-AFP monoclonal antibody was cloned out and designated as MabF22. From isotyping analysis, it was found that monoclonal antibody MabF22 was IgG type with IgG1 heavy chain and k light chain. The binding specificity of MabF22 was analyzed by immunoblotting as well as by ELISA. MabF22 was highly specific, reacting with only AFP-containing samples. The binding affinity was determined by ELISA (free-capture mode) and Scatchard analysis. As a result, the value of Kd was 0.8$\times$10$^{-10}$M. The validity of the MabF22 for AFP assay was examined by two kinds of ELISAs, i.e., non-competitive and competitive ELISA. Both assays revealed that MabF22 reacted well with AFP in sample in a concentration-dependent manner. Standard curve and antibody titration curve were obtained by using purified AFP and MabF22. These results indicate that the monoclonal antibody produced in this study would be useful not only for research purposes but also for further development of immune-diagnostic kit for the measurement of AEP concentration.

• Restorative Dentistry and Endodontics
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• v.26 no.3
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• pp.191-199
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• 2001

목적 - 기질금속단백분해효소(matrix metalloproteinase)는 조직의 염증 및 치유과정에서 숙주세포에서 생성, 분비되어 세포외기질(extracellular matrix)의 분해에 작용한다. 다양한 염증반응에 기질금속단백분해효소가 중요한 역할을 하는것으로 보고되고 있으나 치수 및 치근단 질환에서의 그 역할은 거의 알려져 있지 않은 상태이다. 본 연구에서는 염증이 있는 사람의 치수 및 치근단 조직을 채취하여 Enzymeimmunoassay 및 면역조직화학적 검색을 통해 제1형, 2형, 3형 기질금속단백분해효소의 수준 및 그 분포를 측정하여 치수 및 치근단 병소에서 이 효소의 작용을 알아보는 것을 목적으로 한다. 방법 - 연구재료는 근관치료를 위해 서울대학교 병원 치과 진료부 보존과에 내원한 환자를 대상으로 34개의 치아에서 통상의 근관치료 중 발수한 치수조직과 치근단 수술중 얻은 치근단 병소(n=10)를 이용하였다. 치수는 발수 전에 임상진단을 통해 급성 치수염(n=12), 만성 치수염(n=12), 정상 치수(n=10)로 구분하고 정상치수로 진단된 것을 대조군으로 설정하였다. 채취된 표본은 둘로 나누어 절반은 30분 이내에 5$\mu\textrm{m}$ 두께로 동결절단을 시행하여 조직표본을 제작하였고 deep freezer에 보관하였다가 헤마톡실린-에오신 염색 및 면역조직화학적 검색을 시행하였다. 나머지 조직은 ELISA를 위해 액체 질소에 보관하였다. ELISA를 시행하기전 표본의 단백질 정량을 시행하여 모든 표본의 단백질 양을 50mg/$\mu\textrm{l}$로 일치시키고 Amersham사의 ELISA kit를 사용하여 제1형, 2형, 3형의 기질금속단백분해효소의 양을 측정하였으며 그 결과를 Mann-Whitney U test를 사용하여 각 군간의 통계학적 유의성을 검증하였다. 결과 1. ELISA의 결과 제1형 기질금속단백분해효소의 농도는 모든 실험군에서 대조군보다 유의성있게 높게 나타났다.(p<.05). 또한 급성치수염군의 제1형 기질금속단백분해효소의 농도가 다른 실험군보다 유의성있게 높았다(p<.05). 2. 제2형 기질금속단백분해효소의 경우 급성치수염군과 대조군에서만 유의성있는 차이를 보였다(p<.05). 3. 제3형 기질금속단백분해효소의 경우 급성치수염군에서 대조군이나 만성치수염군보다 유의성 있는 높은 수치를 보였다(p<.05). 4. 면역조직화학검색 결과 염증성 치수에 존재하는 급성 및 만성염증세포 주위로 기질금속단백분해효소에 대한 면역 반응이 존재하였으며 주로 제1형과 제3형 기질금속단백분해효소의 경우 대식세포 및 림파구 주위로 강한 발색제의 침윤양상이 관찰되었다. 5. 치근단병소의 면역조직화학적 검색 결과 만성염증 세포 주변으로 미약한 발색제의 침윤양상이 관찰되었다.

• Proceedings of the KSAR Conference
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• 2004.06a
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• pp.279-279
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• 2004

성장중인 수토끼에서 성성숙 전후의 혈청내 IGF-I 수준과 정자형성의 변화를 12주령부터 28주령까지 2주 간격으로 조사하였다. IGF-I 수준 측정은 non-extraction IGF-I ELISA kit(Biognostic System, Lab, USA)를 이용하였으며 정소내 정자형성은 biopsy needle(22G×3¹/₂ 72㎜×8.89㎝, Becton Dickinson, USA)로 채취한 정소조직을 fled ctyometric ploidy analysis로 DNA histogram을 4개 peak(peak Ⅰ과Ⅱ :1N, peak Ⅱ :2N, peak Ⅳ : 4N, peak Ⅲ과 Ⅳ사이 : S기)로 구분하였다. (중략)