• Environmental Analysis Health and Toxicology
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• 제5권3_4호
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• pp.29-36
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• 1990

Aflatoxins, produced by strains of Aspergillus flavus and Aspergillus parasiticus, can be found worldwide in corn, barley, peanuts, and other commodities. Among this group of toxins, aflatoxin B$_1$was realized to be one of the most potent environmental carcinogens, mutagens and teratogens. It is routinely monitored by methods such as thin layer chromatography, liquid chromatography, fluorodensitometric technique and radioimmunoassay. However, these assays are expensive, necessitate radioactive reagents, and require overnight incubation. In this study, the determination of fungal flora in several sorts cereals has been carried out in order to obtain an appropriate information of the population of fungi. The quantitative analysis of aflatoxin B$_1$has been carried out by High Performance Liquid Chromatography (HPLC) method and Enzyme Linked Immunosorbent Assay (ELISA). The results were summarized as follow: 1) From the 100 samples,313 colonies of fungi were isolated. Among the 313 colonies, 274 were possible to identify into 11 genera. The identified genera were Aspergillus Penicillium, Mucor, Rhizopus, Alternaria, Cladosorium, Fusarium, Circinella, Chrysosporium, Paecilomyces and Phoma. 2) Six of Aspergillus flavus were aflatoxin-producing strains. Aspergillus flavus isolated from sample barleys was contained the highest content (21.8 $\mu\textrm{g}$/ml) of aflatoxin B$_1$. 3) The yield of aflatoxin B$_1$-oxime compound was appromately 75%. Aflatoxin B$_1$-oxime-Human serum albumin was approved by formal consent as complete antigen. 4) Direct competitive ELISA permitted detection of 0.15 ng levels. In the quantitative microanalysis, ELISA was superior to HPLC method.

• 대한수의학회지
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• 제29권4호
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• pp.567-575
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• 1989

This study was conducted to evaluate the possibility of application of a microenzyme-linked immunosorbent assay(micro-ELISA) for the serodiagnosis of specific toxoplasma antibodies in swine sera and this test was performed as a microplate system by coating the polystyrene plates with toxoplasma soluble antigen, incubated serially diluted sera, then added horse radish peroxidase labelled goat anti-swine IgG(r) conjugate followed by o-phenylenediamine as substrate. The color development by enzyme-substrate reaction was determined by the photometric reading [ELISA reader at 490nm (OD)] and visual reading. The soluble antigen was prepared from the tachyzoites in mouse peritoneal cavity. A total of 1,200 swine sera from pig slaughter-house and a total of 116 swine sera from pig breeding station (S-C farm) were tested for the detection of antibodies to Toxoplasma gondii. The results obtained were summarized as follows: 1. The optimal reactions of indirect ELISA for the test sera were determined by the dilution of antigen 1:256 and 1:3,200 of horse radish peroxidase conjugate [anti-swine IgG(r)]. 2. The specific togoplasma antibody(IgG) in pigs infected with Tp artificially were detected as the serum titers of 1:64 or 1:128 at one week postinfection. 3. Of a total of 1,200 swine sera from pig slaughter-house 505 samples of sera were detected as positive (42.1%) and of a total of 116 swine sera from S-C pig breeding station 68 samples of sera as positive (58.6%). 4. The specific antibody(IgG) detection rates against a total of 1,200 test sera from pig slaughter-house were not significant between male (43.1%) and female (40.7%). 5. The indirect ELISA was proved to be a sensitive and specific procedure for the serodiagnosis of swine toxoplasmosis and also evaluated as an effective screening test for the large scale of test samples in laboratory.

• Tuberculosis and Respiratory Diseases
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• 제59권1호
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• pp.47-52
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• 2005

연구배경 : 클라미디아 폐렴균은 임상적으로 중요한 균으로 혈청검사가 진단에 중요하다. 현재 이용되는 micro-immunofluorescence법은 주관적이고 시간이 많이 소요되므로 좀더 객관적으로 진단할 수 있는 ELISA법을 이용하여 만성 기침을 주소로 내원한 환자들의 혈청을 대상으로 검사하고 표준방법과 결과를 비교하였다. 방 법 : 2003년 8월부터 2004년 7월까지 강원대학교병원을 방문한 35명의 성인 환자에서 얻은 혈청을 ELISA법과 micoimmunofluorescence법으로 시행하였다. 비인두 스왑에서 PCR을 시행하였다. 두 방법의 비교는 비모수 상관계수 분석(Spearman)을 하였다. 결 과 : ELISA 검사결과 IgG는 민감도 84.0%, 특이도 60.0%, IgA는 민감도 84.0%, 특이도60.0%, IgM은 민감도 40.0%, 특이도 96.7%의 결과를 보였다. PCR은 3명의 환자에서 양성이었다. 결 론 : ELISA법이 많은 환자들을 대상으로 하는 역학적 연구에서는 좀더 유용하게 이용될 수 있을 것으로 사료된다. 임상적으로 적용되기 위해서는 좀더 연구가 필요할 것으로 사료된다.

• Parasites, Hosts and Diseases
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• 제31권1호
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• pp.49-56
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• 1993

현재 유구낭미충증을 혈청학적으로 진단하는 데에는 낭액항원을 이용한 특이항체검사법으로 micro-ELISA를 널리 이용하고 있다. 이 실험은 효소부착 이차항체를 사용하는 micro-ELISA방법 대신 민감도가 뛰어난 것으로 알려진 ABC-ELISA나 Protein-ELISA로 바꾸어 사용하면 검사의 민감도를 보다 개선할 수 있는지를 알기 위하여 실시하였다. ABC-ELISA에 의한 항체검사는 낭액항원 단백질 $2.5{\;}\mu\textrm{g}/ml$, 혈청 희석 1:10,000, biotinylated no-hmn IgG 1:100,000 회석, peroxidase conjugated streptavidin 1:40,000 희석 및 2,2-azino-di(3-ethylbenztlllazoline) sulfnnlc acid발색제를 사용하는 조건으로 실시하였고 415 nm에서 흡광도를 측정하였다. Protein A-ELISA는 항원단백질 $2.5{\;}\mu\textrm{g}/ml$, 혈청은 1:200 회석, HRP-Protein A 1:20,000 희석 및 ABTS 발색제를 사용하는 조건으로 실시하고 415 nm에서 흡광도를 측정하였다. 유구낭미충증 환자 115 명의 혈청을 검사한 바 민감도는 micro-ELISA 81.7%, Protein A-ELISA 82.6%, ABC-ELISA 86. 1%이었다. 다른 기생충성 질환자, 비기생충성 질환자 및 대조군 등 165명 혈청에서 특이도는 각각 88.5%, 93.3%, 93.8%이었다 세 가지 ELISA방법에 의한 항체가 사이에는 상관계수 0.84~0.86의 높은 상관관계가 성립하였다. 이상의 결과 ABC-ELISA는 micro-ELISA에 비하여 민감한 혈청학적 방법이고 실제 유구낭미충 특이항체 검사상의 특이도에서도 차이가 있다는 것을 알 수 있었다. 따라서 ABC-ELISA는 유구낭미충증의 혈청학적 진단에서 혈청 및 뇌척수액 등 검체를 적게 사용할 수 있다는 장점이 있다는 것을 알 수 있었다.

• 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.

• Journal of Biosystems Engineering
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• 제32권6호
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• pp.440-447
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• 2007

In this study, a biosensor was developed using an enzyme-linked immunosorbent assay (ELISA) to rapidly measure the fungicide iprovalicarb residues in agricultural products. The biosensor was designed to include micro-pumps and solenoid valves for fluid transport, a spectrophotometer cuvet as a reaction chamber, a photodiode with a light-emitting diode for optical density measurement, and a control microcomputer to implement assay. The rate of change in optical density of the cuvet was read as final signal output. Micro-pumps were evaluated to investigate their delivery capability, the highest values of the error and the coefficient of variation were 4.3% and 4.6% respectively. As the incubation period was reduced from 15 minutes to 11 minutes to shorten the total processing time, the sensor sensitivity was decreased as the antibody dilution ratio was reduced to a half. The maximum usable period of the coated cuvet was found to be two days with 1% error limit. To predict the concentration of the iprovalicarb residue in agricultural products, a linear calibration model was obtained with r-square values of 0.992 for potato and 0.985 for onion. In validation test for the samples of potatoes and onions against the high performance liquid chromatography, very high correlation values were obtained as 0.996 and 0.993 respectively. Using the cuvet immobilized with antigen, it took 21-minutes for the biosensor to complete the measuring process of the iprovalicarb residues.

• Parasites, Hosts and Diseases
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• 제23권1호
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• pp.95-101
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• 1985

Fascioliasis in cattle is one of the most common and very serious trematode diseases in Korea. In the present study, the enzyme linked immunosorbent assay (ELISA) was applied in the diagnosis of fascioliasis using antigen of Fasciela hepatica, perokidase of conjugate anti-cattle Is G and orthophenylenediamine as a substrate by micro-method technique of Volley et at. (1976b) and MacLaren (1978) with a slight modification. Results obtained from the present study are as follows. 1. In assay for optimal dilution of stock antigen, the antigen (protein contents; 0. Bmgymz) was diluted from 1150 to 1/600 with carbonate buffer (pH 9.6), and then absorbance values were measured with 1/100 diluted sera. The regression equations between the OD values of ELISA and dilution of antigen were log Y: -0.181-0.00127X in infected sera, and log Y: -0.578-0. 000879X in normal sera. The significantly higher (p<0.05) OD value was observed in the former. 2. In assay for optimal dilution of sera, the sera were diluted from 1125 to 1/400 with in PBSJ Tween 20 (pH 7.4), and absorbance values were measured with 1/200 diluted antigen. The regression equation between the OD values of ELISA and dilution of sera were log Y: -0.1540-0.0007238X in infected sera and log Y: -0.4834-0.00116X in normal sera. The former was higher than the latter (p<0.05). 3. In the 27 cases of negative intradermal test, OD values of the ELISA are $0.447{\pm}0.144$, the 95% confidence interval (Mean+2 H SD) of the values was 0.735, and there was no case over the values. Therefore, the sensitivity of the antigen to diagnose fascioliasis was 100% in the negative case. The OD value 0.7 which is designed as a criterion (detection level of positive one) is useful for the performance of the ELISA in fascioliasis. 4. According to the OD value of criterion in the regression equations, the optimal dilutions of stock antigen and serum were 1/250 and 1/100, respectively. 5. In the 58 cases of fascioliasis from which the adult could be found in the bile ducts, the OD value was $0.846{\pm}0.224$. The 75% (44 cattle) among them had higher value with compared to the criterion, and the 60% (20 cattle) of the cases of proliferative cholangitis of 33 cattle which had been infected previousely with Fasciola sp. is higher than the criterion. 6. Prevalence of fascioliasis was 43.4% in the application of the ELISA to 272 cattle which were reared in Jeonbug district.

• Journal of Veterinary Science
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• 제24권2호
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• pp.20.1-20.13
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• 2023

Background: As Actinobacillus pleuropneumonniae (APP) infection causes considerable losses in the pig industry, there is a growing need to develop effective therapeutic interventions that leverage host immune defense mechanisms to combat these pathogens. Objectives: To demonstrate the role of microRNA (miR)-127 in controlling bacterial infection against APP. Moreover, to investigate a signaling pathway in macrophages that controls the production of anti-microbial peptides. Methods: Firstly, we evaluated the effect of miR-127 on APP-infected pigs by cell count/enzyme-linked immunosorbent assay (ELISA). Then the impact of miR-127 on immune cells was detected. The cytokines tumor necrosis factor (TNF)-α and interleukin (IL)-6 were evaluated by ELISA. The expression of cytokines (anti-microbial peptides [AMPs]) was assessed using quantitative polymerase chain reaction. The expression level of IL-6, TNF-α and p-P65 were analyzed by western blot. The expression of p65 in the immune cells was investigated by immunofluorescence. Results: miR-127 showed a protective effect on APP-infected macrophage. Moreover, the protective effect might depend on its regulation of macrophage bactericidal activity and the generation of IL-22, IL-17 and AMPs by targeting sphingosine-1-phosphate receptor3 (SIPR3), the element involved in the Toll-like receptor (TLR) cascades. Conclusions: Together, we identify that miR-127 is a regulator of S1PR3 and then regulates TLR/nuclear factor-κB signaling in macrophages with anti-bacterial acticity, and it might be a potential target for treating inflammatory diseases caused by APP.

• Parasites, Hosts and Diseases
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• 제33권4호
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• pp.395-398
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• 1995

강원도 홍천에 거주하는 56세 구부가 갑자기 우상부 복통 고열 오한을 주소로 내원하였고 복 꾸 초음파검사와 콤퓨터단층촬영 결과 2-3 cm 직경의 불규칙한 꼴의 동공이 간 우엽에서 여러개 관찰되었다. 간생검상 샤르코-라이덴 결정을 포함한 심한 중심성 괴사 주위로 조직구. 호산구등 염 증세포가 침윤되어 주위 조직과 명확히 구분되었다. 효소면역측정법을 이총한 간질 특이 항체 혈청 걸사에서 양성이었다. 간 병변은 프라지관텔 투약 6개웍 후에 모두 사라졌으나. 특이 항체가와 호산구 증가는 투약 6개월 16개월 후에도 정상화 되지 않았다. 방사선학적 소견 조직병리학 소견 및 혈청학 검사 결과를 종합할 때. 초기 침습성 간질증으로 진단하였으며 프라지콴텔에 의하여 치료되지 않은 예이었다.

• Journal of Chest Surgery
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• 제38권12호
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• pp.807-814
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• 2005

배경: 체외순환은 심정지를 필요로 하는 모든 심장 수술에서 정지된 심장의 기능을 대신하여 환자의 말초 장기의 혈액순환을 유지하기 위한 필수적인 과정이다. 그러나 체외순환은 필연적으로 인공도관을 관류하는 특성상 신체의 혈관계를 흐르는 혈류와 달리 혈액손상에 따른 전신성 염증반응을 피할 수 없으며, 이러한 전신성 염증반응과 함께 말초혈관의 미세혈관 순환장애가 체외순환동안에 원발장기의 손상을 초래하는 것으로 생각된다. 저자들은 전신성 염증반응을 일으키는 주된 혈액성분인 백혈구를 제거한 충진액을 사용하여 전신성 염증반응을 줄일 수 있는가를 확인하고, 체외순환도중 위점막의 산도를 측정함으로써 위점막의 미세혈류에 대한 백혈구 제거 충진액의 효과를 확인하기 위하여 실험을 진행하였다. 대상 및 방법: 실험군은 15마리의 한국산 잡견을 충진액의 성분에 따라 비혈액성 충진액군, 백혈구 제거 혈액성 충진액군, 백혈구 비제거 혈액성 충진액군으로 각각 5마리씩 세 군으로 나누었다. 세 군 모두에서 2시간의 체외순환 및 연속된 4시간의 마취유지를 시행하였으며, 체외순환 전과 체외순환 후 1시간, 2시간, 체외순환 종료 후 2시간 4시간에 위점막 이산화탄소 농도와 산도, 동맥혈 이산화탄소 분압 과 호기말 이산화탄소 분압을 측정하고, 염증반응의 지표검사를 위하여 동맥혈을 채혈하였다. 전신성 염증반응의 정도는 채취한 동맥혈에서 ELISA (enzyme linked immunosorbent assay)법을 이용하여 IL-8의 수준을 검사하였다. 결과: 1, 위점막의 이산화탄소 농도는 백혈구 제거 혈액성 충진액군이 백혈구 비제거 혈액성 충진액군과 비혈액성 충진액군에 비하여 유의하게 낮았다(p=0.02, 0.01). 2. 위점막의 산도는 백혈구 제거 혈액성 충진액군과 백혈구 비제거 혈액성 충진액군간에 유의한 차이를 보였다(p=0.01). 3. 전신성 염증반응의 정도를 확인하기 위하여 측정한 IL-8의 수준은 백혈구 제거 혈액성 충진액군과 비혈액성 충진액군이 백혈구 비제거 혈액성 충진액군에 비하여 유의하게 낮았다(p=0.01, 0.01). 결론: 백혈구를 제거한 혈액성 충진액을 사용하는 것이 체외순환중 위점막의 미세순환 장애를 방지하고 전신성 염증반응을 감소시킬 수 있음을 확인하였다.