• The Plant Pathology Journal
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• 제36권5호
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• pp.509-514
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• 2020

Satsuma dwarf virus (SDV) seriously damages citrus production by reducing the quality and yield of fruit. To avoid contamination with SDV, mother trees are checked to be SDV-free in advance of nursery tree distribution. In this study, we compared an immunochromatographic assay (ICA) kit with double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) for large-scale diagnosis of SDV in orchardgrown trees in Shizuoka Prefecture, Japan. The two methods gave conflicting results for 11 of 1,705 samples, all of which were negative by DAS-ELISA but positive by ICA. The samples scored as positive by either DAS-ELISA or ICA were analyzed by reverse transcription polymerase chain reaction and all were confirmed to be positive. These results validate the use of ICA as a screening method for large-scale diagnosis. Strain discrimination revealed that 16 of 22 isolates belonged to SDV, while citrus mosaic virus (CiMV) infection only and co-infection (SDV and CiMV) were in a minority.

• 대한수의학회지
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• 제49권1호
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• pp.39-44
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• 2009

Transmission of avian viruses both bird-to-bird and from birds to non-avian species is a major health concern. Newcastle disease virus (NDV) is an economically important avian virus that poses substantial risks to the poultry industry. Rapid and sensitive diagnostic methods, such as the enzymelinked immunosorbent assay (ELISA), are required to track such infections. To develop an ELISA for detecting anti-NDV antibody in avian sera, the nucleocapsid protein (NCP) gene of the NDV La Sota strain was cloned and expressed in Escherichia coli and the 513-amino acid recombinant NCP was purified by Ni-NTA affinity chromatography. To evaluate its ability to replace NDV whole virus antigen as a coating antigen, NCP-coated and whole NDV-coated ELISAs were tested and compared using a panel of NDV positive antisera from chickens. Results using purified NCP were highly correlated with those obtained using whole NDV (r= 0.927), demonstrating that recombinant NCP expressed in Escherichia coli is a suitable substitute antigen for whole NDV in a diagnostic ELISA.

• 한국식품영양과학회지
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• 제30권4호
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• pp.600-604
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• 2001

감마선 조사된 새우의 신속한 판별을 위한 방법으로 갈색새우의 TPM을 항원으로 하여 개별적으로 생산된 M-IgG와 P-IgG를 이용한 Sandwich ELISA를 분석법으로 확립하였다. M-IgG를 항원 포획을 이한 coating 항체로 사용하고, P-IgG를 포획된 TPM에 대한 반응항체로 사용하였을 때, 12.5에서 50$\mu\textrm{g}$/mL의 농도 범위에서 TPM을 정량할 수 있었다. 감마선 조사된 새우의 TPM 농도는 선량에 의존하며 감소하였고, 감마선 조사와 가열 또는 냉동 등의 병용 처리에서도 선량에 의존하며 감소하였다. 이 결과는 면역분석기법의 하나인 Sandwich ELISA가 감마선 조사된 새우의 검지법으로 이용될 수 있다는 가능성을 나타내었다.

• Biomolecules & Therapeutics
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• 제18권2호
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• pp.184-190
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• 2010

Clinical cases of pure red cell aplasia (PRCA) have been reported during the recombinant human erythropoietin (EPO) therapy for the anemia patients. PRCA is a rare hematological disorder leading to a severe anemia due to an almost complete stop of red blood cell production. Antibody (Ab)-associated PRCA is caused by the EPO-neutralizing Abs that eliminate the biological activity of EPO. In order to detect anti-EPO Abs in human sera, we performed conventional ELISA, directly coated bridging ELISA, and streptavidin coated bridging ELISA, and compared their sensitivity and specificity. Some false positive results were obtained in the conventional ELISA. One positive sample was detected successfully by streptavidin coated bridging ELISA, which was not appeared in the directly coated bridging ELISA. In conclusion, streptavidin coated bridging ELISA was substantially sensitive and specific format and one out of sixty-eight serum samples was proved to be anti-EPO positive.

• Current Research on Agriculture and Life Sciences
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• 제1권
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• pp.195-199
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• 1983

우백혈청(牛白血病)virus에 대한 혈청항체의 측정(測定)을 위한 간접효소면역법(間接酵素免疫法)(ELISA)의 확립(確立) 및 ELISA법(法)의 감도(感度)를 알기 위하여 한천(寒天) gel 면역확산법(免疫擴散法)(ID)과 비교검토(比較檢討)하였으며 한우(韓牛), 유우(乳牛) 및 육우(肉牛)등 264두(頭)의 혈청(血淸)을 공시(供試)하였다. 혈청(血淸)을 가(加)하지 않은 공(孔)의 흡광치(吸光値)(C)로서 혈청치(血淸値)를 제(除)한 후, 표준(標準)BLV항체흡성혈청치(N)로서 피검혈청치(被檢血淸値)(T)를 나눈치(値)(T-C/N-C)가 1.5이상(以上)인 것을 BLV항체 양성(陽性)으로 하였을 때 gp-ID의 성적(成績)과 98.5%(259/263)가 일치(一致)되었다. gp-ID 향성혈청(陽性血淸) 145예(例)중 144예(例)가 ELISA 양성(陽性)으로 99.6%, gp-ID 음성혈청(陰性血淸) 118예(例)중 115예(例)가 ELISA음성(陰性)으로 97.5%가 일치(一致)되었다. 이상에서와 같이 gp-ID용(用)의 BLV 항원(抗原)을 사용한 ELISA법(法)을 gp-ID와 동등(同等) 또는 더 감도(感度)가 우수한 BLV의 혈청항체측정법 임이 입증되었으며 실용성(實用性)이 충분(充分)하다고 사료(思料)된다.

• 한국식품영양과학회지
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• 제27권6호
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• pp.1152-1159
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• 1998

In order to develop more rapid and reproducible analysis of biotin known as vitamin H, attempts were made to establish the condition for enzyme linked immunosorbent assay(ELISA) compared with traditional microbiological assay(MBA). Antibiotin and antiserum were obtained from the immunized rabbits injected with emulsion of biotin KLH conjugate and Freund's adjuvant. The antiserum showed cross reactivity on biocytin, a derivative of biotin, which is converted to biotin in intestine, at the rate of 177%(median inhibitory concentration(IC50)=12.58ppb), but not on other derivatives such as desthiobiotin, diaminobiotin and 2 imino biotin. Specific antibody for biotin was purified from the antiserum through protein A column and desalting column. The conditions of competitive direct ELISA (cdELISA) were established. Detection range of biotin concentration by cdELISA was 0.01∼300ng/ ml(ppb). In the spike test with milk, fruit flake and pine carrot juice, the correlation coefficient between two methods of MBA and ELISA was reliably consistent at the value of r=0.992. But detection of biotin by microbiological assay(MBA) was rather restricted in range and nonspecific. Detection range of biotin by MBA was 0.1∼0.5ng/ml(ppb). It showed cross reactivities on biocytin and desthiobiotin at the rate of 80.1% and 66.7%, respectively. In conclusion, ELISA revealed a significant improvement compared with MBA for the biotin detection in terms of sensitivity, detection range and cross reactivity. In addition, a variety of samples could be analyzed rapidly and conveniently at one time by using ELISA. These results strongly suggest that the ELISA is very promising for the practical application to detect biotin contents in a wide range of food stuffs.

• 대한수의학회지
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• 제38권4호
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• pp.793-802
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• 1998

Salmonellosis caused by a number of serotypes of Salmonella is an infectious, acute or chronic, zoonotic disease and characterized by enteritis and diarrhea, septicemia in animal. In these studies we investigated the prevalent serotypes of Salmonella causing animal salmonellosis in Korea and the 71 strains of Salmonella spp. were isolated from materials such as mesenteric lymph nodes, fecal samples from slaughtered animal. With the identification test results, the most prevalent serotypes were, in order, S stanley 31 strains (43.7%), S typhimurium 19 strains (26.8%) and S montevideo 11 strains (15.5%), respectively. And we could establish the method for detection of antibodies to broad variety of Salmonella serotypes. Lipopolysaccharide(LPS) antigen extracted from Salmonella was more sensitive and specific than outer membrane protein antigen from that for detection of Salmonella antibody by using an indirect ELISA. The optimal concentration of antigen was 100ng/ml of LPS, the dilutions of conjugate and serum were 1 : 1,000~2,000 and 1 : 200~400, respectively. The mix LPS-ELISA which was used by mixing LPS from S typhimurium (group B), S choleraesuis (group C) and S enteritidis (group D) were more rapid and effective than that used LPS from individual strain for detection of Salmonella serogroup O4, O7 and O9 antibody at the same time. We could obtain the high values of optical density ($0.73{\pm}0.32$) by mix LPS-ELISA on the farm which had occurred salmonellosis, but very low values of $0.17{\pm}0.06$ on the negative farm of salmonellosis. So, the mix LPS-ELISA may be used to monitor the serological surveillance for the presence of infection with a number of serotypes of Salmonella and would be useful for prevention and control of salmonellosis in animal.

• 한국가금학회지
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• 제45권1호
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• pp.17-28
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• 2018

전염성기관지염은 전세계적으로 양계산업에 문제시되고 있는 질병으로서 이를 예방하기 위한 연구가 활발히 진행되고 있다(Dhama et al., 2014). 본 연구는 전염성기관지염 백신을 진행한 여러 계군의 혈청을 수집해 국내회사와 다국적기업에서 제조한 ELISA 키트의 검출능 차이를 확인하고자 하였다. 육용원종계부터 산란계까지 총 35개의 계군으로부터 주기적인 채혈을 통해 얻어진 혈청을 사용해 두 가지 키트의 검출능 비교시험을 진행한 결과, 국내회사에서 제조된 ELISA 키트가 더 높은 항체가를 나타냈으며, 이는 미량의 항체도 국내회사 제조 키트를 사용하게 되면 더욱 민감하게 검출될 수 있으며, 농장의 상황 판단에 더욱 도움이 될 수 있을 것임을 시사한다. 이러한 결과를 토대로 비추어 보았을 때, 국내에서 발생하고 있는 전염성기관지염에 맞춰 제작된 국내회사 제조 키트가 더 검출능이 뛰어난 것으로 판단된다. 또한, 주기적인 혈청을 이용한 항체가 검사 결과 전염성기관지염 백신 후 항체가 변화 추이를 관찰할 수 있었다.

• 한국식품위생안전성학회지
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• 제32권1호
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• pp.75-81
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• 2017

본 연구에서는 고등어 어육을 고감도로 보다 신속하게 검출하기 위하여 고등어 어육 중 TSSP를 이용해 단클론성 항체를 개발하고 이를 이용하여 간접 효소면역분석법(iELISA)과 western blot의 검출한계를 확인하였다. 먼저 비 열처리와 열처리를 한 고등어 어육 중 존재하는 TSSP를 확인하고, 단백질 추출에 주로 사용되는 버퍼의 종류에 따른 추출법 효율을 확인하기 위하여 전기영동과 단백질 정량을 실시하였다. 그 결과 열처리한 추출물은 비 열처리한 추출물보다 TSSP가 2배 정도 많이 추출되었으며, TSSP로 추측되는 37 kDa 부근에 형성된 밴드의 선명함과 굵기를 육안으로 비교한 결과 carbonate buffer로 추출하였을 때 밴드가 가장 두드러지게 형성되었고, 단백질 정량 결과에서도 carbonate buffer를 이용한 추출물의 단백질 농도가 가장 높은 것을 확인하였다. 이후, 생 고등어 어육을 열처리법으로 추출하여 항원을 준비하고 6주령 BALB/c mouse에 면역한 후 세포융합 및 클로닝을 통해 3A5-1번, 2번, 9번 및 16번 4종의 hybridoma cell을 확보하였다. 이렇게 개발된 항체는 수산물, 축산물, 농산물과의 교차반응성확인을 통하여 고등어 단백질에만 특이성을 가지는 것을 확인하였다. iELISA와 western blot법의 검출한계를 확인한 결과 고등어가 1% 첨가된 수준까지 검출할 수 있으며, 특히 3A5-2와 3A5-9번 항체는 1%에서도 높은 흡광도를 나타내어 민감도가 매우 높은 항체로 확인되었다. 따라서, 개발된 고등어 TSSP 특이항체를 이용한 iELISA법과 western blot법은 가공품에 혼입될 수 있는 식품 알레르겐인 고등어를 보다 신속하고 민감하게 분석할 수 있는 분석 도구로서 활용이 가능할 것으로 판단되며, 개발된 항체는 고감도 바이오센서 개발에 충분히 활용이 가능한 바이오인자로 확인되었다.

• 생명과학회지
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• 제9권4호
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• pp.453-459
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• 1999

Baculovirus(WSBV) was isolated from infected Penaeide was collected from shrimp farm at southern sea of Korea from 1993 to 1995. The Infectious virus was purified and used for diagnosis of infected shrimp. Anti-viral serum were used for immunological detection as enzyme linked immunoabsorbent assay(ELISA) and indirect fluorescent antibody technique(IFAT). In IFAT, stomach, lymphoid organ and antenae gland of infected shrimp showed fluorescent reaction. In ELISA, tissues of spontaneously infected shrimp appeared higher O.D. values than in artificial infected shrimp. Primer set was constructed from sequence of 420bp of cloned Baculovirus(WSBV) genome. Specific band for infected shrimp was detected in Polymerase chain reaction(PCR)