• Journal of Microbiology and Biotechnology
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• 제9권6호
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• pp.764-772
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• 1999

To develop an enzyme-linked immunosorbent assay (ELISA) for detecting mold, we produced anti-mold polyclonal antibodies by immunizing extracellular polysaccharide (EPS) of Aspergillus flavus or Penicillium citrinum in rabbits subcutaneously. Using the purified antibody (Ab) and Ab-HRP conjugate, a sandwich ELISA for EPS was established. The standard curve of the ELISA showed the detection limit for P citrinum EPS to be $0.003{\;}\mu\textrm{g}/ml$. The cross-reactivities of the anti-P citrinum EPS Ab toward components of P citrinum such as EPS, liquid, and solid culture mycelium were 100, 10.5, and 0.58%, respectively, and those toward components of A. flavus such as EPS, liquid and solid culture mycelium, and spore were 300, 0.67, 0.29, and 0%, respectively. When the reactivities toward culture broths of 59 mold strains were tested by the sandwich ELISA, most of the Aspergillus (16 of 18) and Penicillium (14 of 16) strains along with one of the two Cladosporium strains gave positive signals in the culture broths even when diluted 1,000 fold, while the rest of species such as Fusarium, Absidia, Alternaria, and Candida gave negative signals. When the water extracts of 30 corn samples were analyzed by the sandwich ELISA, the EPS in the com could be detected in the concentration range of $0.1-1.6{\;}\mu\textrm{g}/g$.

• Journal of Microbiology and Biotechnology
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• 제14권4호
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• pp.686-692
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• 2004

To develop the enzyme-linked immunosorbent assay (ELISA) for the analysis of N-acetylehitooligosaccharides (NACOS) and chitooligosaccharides (COS), specific antibodies (Abs) were produced, and their properties were compared. N-acetylehitohexaose (NACOS6), chitohexaose (COS6), and COS mixture (COSM) conjugated to bovine serum albumin (BSA) were used to immunize rabbits. By the use of specific Abs and NACOS6-horseradish peroxidase (HRP), COS6-HRP, and COSM-HRP conjugates, competitive direct ELISA (cdELISA) was developed. The detection limits of NACOS6 by the anti-NACOS6 Ab and COS6 by the anti-COS6 and the anti-COSM Abs in the cdELISAs were about 0.2, 2, ana 2 ng/ml (ppb), respectively. In the cdELISA, the anti-NACOS6 Ab was found to recognize NACOS3-NACOS6, but not N-acetyl-D-glucosamine (GlcNAc), NACOS2, and COSs; the anti-COS6 Ab recognized COS2-COS6 and COSM, but not glucosamine (GlcN) and NACOSs. The recognition pattern of the anti-COSM Ab was almost the same as that of the anti- COS6 Ab, except that the former recognized COS2 and COS3 slightly better than the latter.

• 식물병연구
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• 제16권2호
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• pp.121-124
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• 2010

박과작물에 발생하는 종자전염 바이러스 3종(CGMMV, KGMMV, ZGMMV)에 대한 검출을 위해 IC-RT-PCR의 적용 가능성을 시험하였다. IC-RT-PCR을 이용하여 감염 종자와 잎으로 부터 바이러스의 특이적인 검출이 가능하였으며, 검출 민감도는 ELISA 보다 100배 이상 높았다. 또한 반응을 마친 ELISA 마이크로플레이트에 남아있는 항원을 IC-RT-PCR의 template로 사용할 경우 ELISA 결과를 곧바로 확인할 수 있었다. 따라서 IC-RT-PCR에 의한 본 검정방법은 박과작물의 대규모 포장검사 및 종자검사에 편리하게 사용될 수 있을 것으로 기대된다.

• 대한수의학회지
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• 제49권2호
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• pp.121-125
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• 2009

Paratuberculosis caused by Mycobacterium avium subspecies paratuberculosis (Mpt) is a chronic infectious enteric disease with deleterious impact on the performance in ruminants. In Korea, ELISA has been introduced to detect antibodies to Mpt in individual cattle. However, comparison study with ELISA has not been studied until now. In total, a panel of 899 serum samples obtained from dairy cattle was analyzed with two commercial ELISAs for Mpt to assess the performance. Two ELISAs employed in this study were both licensed worldwide. Two ELISAs applied onto same serum samples showed the moderate agreement (kappa value = 0.60). There was non-significant McNemar test (p = 0.0614) between two ELISA results indicating that each proportion detected by two kits did not differ. In addition, the percent agreement between two ELISA results was turned out to be 96.8% which interpreted excellent reproducibility. It was shown from this study that two ELISAs revealed moderate kappa agreement performance. The implication raised is that when ELISAs as diagnostics are used to detect Mpt in individual cattle, positive reaction by either ELISA should be interpreted as serologically Mpt positive due to presumed low sensitivity of ELISAs and their test agreement being less than 100%.

• 약학회지
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• 제41권1호
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• pp.74-80
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• 1997

A polygonal antibody against recombinant hirudin was raised for the development of a ELISA in biological fluids. Recombinant hirudin was conjugated to maleimide activated carrie r protein, KLH and injected to a rabbit. The third booster collection of antiserum was used as primary antibody for the ELISA. The titer for the detection antibody was determined. The direct ELISA could determine the concentration of hirudin in the range of ~10ng/ml. Affinity pulified IgG was obtained and conjugated to horseradish peroxidase. Purified IgG and IgG-HRP could be used as capture and detection antibody, respectively. Although sandwich ELISA would not give the satisfactory results. it could apply for the detection of hirudin level in the range of ~20 ${\mu}$g/ml.

• 약학회지
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• 제55권1호
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• pp.16-21
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• 2011

HER-2 (Human Epidermal Growth Factor Receptor-2) is a protein giving higher aggressiveness in human breast cancers. Trastuzumab is a monoclonal antibody that targets HER-2 and is known to extend survival across all stages of HER2-positive breast cancer. In this study, we attempted to development of a quantitative ELISA (Enzyme-Linked ImmunoSorbent Assay) for evaluating anti HER-2 antibodies using human HER-2 recombinant proteins to support antibody producing processes and pharmacokinetic studies. We established direct or indirect ELISA method for the trastuzumab-like protein combined human recombinant HER-2. The ELISA method will prove to be great value in quantitating anti-HER-2 antibodies levels for developing anticancer antibodies.

• 한국군사과학기술학회지
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• 제13권3호
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• pp.478-485
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• 2010

The vaccine is biological pretreatment that improves immunity to a particular disease. We can get immunity from producing antibody with injection antigen which has ability to defense against the disease. The ELISA is the most widely used method to measure antibody titer. We have developed and performed validation of ELISA according to the guideline of KFDA and ICH. In this paper, we have verified ELISA method is an excellent method to measure the titer of anti-PA antibody. We have constructed recombinant protective antigen among anthrax toxins and used as antigen of ELISA. In this validation, we have evaluated precision (repeatability, interlaboratory precision), specificity, linearity(range) and LOD, which are validation articles suggested by guideline. Inter-person precision was replaced with inter-laboratory precision. From the results, we have confirmed high precision in all experiments with CV under 20%.

• 한국미생물·생명공학회지
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• 제24권5호
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• pp.630-635
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• 1996

For a survey of the occurrence of aflatoxin M$_{1}$ (AFM$_{1}$) in domestic cow's milk, we developed an enzyme-linked immunosorbent assay (ELISA) system, and quantitated the toxin in cow's milk. In order to produce specific antibodies AFM, conjugated to bovine serum albumin (AFM$_{1}$-BSA) and Freund's adjuvant were immunized subcutaneously to rabbits. By use of the antiserum showing the highest titer and AFB$_{1}$-HRP conjugate, we established a competitive direct ELISA (cdELISA) for AFM$_{1}$, whose detection limit was 0.003 ppb. The cross-reactivities of the antiserum against aflatoxin M$_{1}$ M$_{2}$, B$_{1}$, B$_{2}$, G$_{1}$, G$_{2}$, B$_{2a}$, and G$_{2a}$, were 100, 29.9, 25.0, 2.7, 13.0, 0.65, 0, and 0%, respectively. When the cdELISA was applied to the cow's milk spiked with AFM$_{1}$ and followed by cleanup with C$_{18}$ cartridge, the mean recovery of the assay was 104% (mean of CV, 6.4%) in the final concentration of 0.01-1 ppb (10-1, 000 ppt). When cow's milk samples gathered from markets and farms were assayed by the cdELISA, the mean concentration and SD of AFM$_{1}$ was 80.4 $\pm$ 55.0 ppt (n=64; range, 5.6-280 ppt).

• 한국식품위생안전성학회지
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• 제29권2호
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• pp.158-163
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• 2014

본 연구에서는 상용화 되고 있는 PCR 및 ELISA kit를 사용하여 국내에서 유통되고 있는 소, 돼지, 닭, 오리, 칠면조, 염소, 양, 말 등 8종의 식육, 혼합육, 그리고 식육가공품에 대하여 축종 감별능력을 평가하였다. 신선육에 대한 RAW meat ELISA kit$^{(R)}$의 검출한계는 축종별 함유율 0.20%~0.05% 이었고, 열처리 혼합육에서는 열처리 온도 및 시간, 그리고 축종별로 검출한계는 함유율 1.0%~0.05% 이하까지 다양한 차이를 나타내었다. 8종의 식육에 대한 축종별 감별력은 소 94.5%, 돼지 93.3%, 양 90.0%, 오리, 염소, 말, 칠면조 모두에서 100%를 나타내었다. Powercheck Animal Species ID PCR kit$^{TM}$의 경우에는 함유율 0.05%의 검출한계를 나타내었고 8종의 모든 축종에서 100%의 특이도를 나타내어 축종별 감별력이 우수한 것으로 나타났다. 또한, 햄, 소시지, 분쇄가공품, 식육추출가공품 등 총 60개 식육가공품에 대한 Cooked meat ELISA kit$^{(R)}$의 감별력은 햄(35.3%), 소시지(13.6%), 분쇄가공육(12.5%)의 순으로 나타났으며, 2종 이상의 혼합육에서는 상대적으로 낮은 감별력을 보여 제조과정에서 식육간 교차오염에 의한 혼입가능성이 있는 것으로 확인되었다. 쇠고기 육포 54개 제품에 대하여 다른 고기 혼입여부를 PCR Kit로 검사한 결과 13개 제품에서 돼지고기 유전자가 검출되었지만 ELISA Kit에서는 모두 음성으로 나타났다. PCR 양성 시료의 제조공정 중 교차오염 여부를 조사한 결과, 텀블러, 채반, 절단기, 건조기가 쇠고기 및 돼지고기 육포 생산라인에 동일하게 사용되어 교차오염에 의한 혼입으로 추정되었다. 종교적 이유 및 일부 특정 육류에 대한 알러지 반응 등 식품안전 확보차원에서 제품의 원재료의 올바른 표시와 식육간 교차오염이 발생되지 않도록 철저한 품질관리가 되어야 할 것으로 판단된다.

• Parasites, Hosts and Diseases
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• 제55권3호
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• pp.247-254
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• 2017

ELISA has been used for the diagnosis of toxoplasmosis, but it is being gradually replaced by a rapid diagnostic test (RDT). We compared and analyzed ELISA and RDT results using the sera collected during 4 consecutive years from residents of Gyodong-do (Island), Incheon-city, Korea. Sera from 921, 993, 940, and 838 adult residents were collected on a yearly basis (2010-2013). ELISA was performed by using a crude extract of T. gondii RH strain antigen and IgG/IgM RDT mounted with recombinant fragment of major surface antigen (SAG1), GST-linker-SAG1A, were applied to the sera. Comparison between groups was analyzed by the Student's t-test. The positive seroprevalence surged from 14.7% (135/921, 2010), 23.1% (231/993, 2011), 23.6% (222/940, 2012), and 32.1% (269/838, 2013) by ELISA. In contrast, RDT showed a more moderate increasing trend from 21.7% (200/921, 2010), 25.5% (253/993, 2011), 28.9% (272/940, 2012) and 33.1% (277/838, 2013). Discrepancies between ELISA and RDT were noted near the cut-off value. At the OD 0.15-0.24 range, RDT could detect 16.1% (169/1051) more positives, which suggests an early or acute toxoplasmosis, but at the OD 0.25-0.34 range, ELISA could detect 35.9% (92/256) more positives of possible chronic infections. Over the OD > 0.35 ELISA and RDT agreed in the majority of the cases. This surge in seroprevalence may be caused by the organic agriculture in addition to eating behavior or increase in pets among Koreans. These facts may be applied on a full-scale national survey using RDT to supplement ELISA to define the characteristics of the infection.