• 한국미생물·생명공학회지
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• 제25권4호
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• pp.414-419
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• 1997

In order to develop an enzyme-linked immunosorbent assay (ELISA) for deoxynivalenol (DON) in com, we produced a specific monocl- onal antibody and established ELISA conditions. After the spleen cells from mice immunized with DON-bovine serum albumin conjugate were fused with S$_{p}$2/0 myeloma cells, a hybridoma cell 3G7 producing anti-DON antibody was screened by ELISA. From the standard curve of competitive direct ELISA (cdELISA) using 3G7 monoclonal antibody and DON-HRP conjugate, the detection range of DON showed 3-3,000 ng/ml (ppb). The monoclonal antibody showed some cross-reactivities against DON analogues such as 15 acetyl-DON (110%), nivalenol (5.0%), 3 acetyl-DON (1.7%), fusarenon-x (0.72%), and T-2 (0.59%). When the cdELISA was applied to the spiked coms after extracting with 60% methanol and diluting 5- fold with washing buffer, the assay recoveries of DON were 313, 163, 106, and 88.9% (av., 168%) in the levels of 200, 600, 2,000, and 6,000ng/g, respectively. For the quantitation of DON in coms, 30 samples kept under two different storage conditions of cold and room temperature were assayed by cdELISA. The mean detection concentrations were 595 (detection range, 0-2,750) and 2,448 (detection range, 0-4,500) ppb, respectively.

• Journal of Microbiology and Biotechnology
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• 제14권2호
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• pp.385-389
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• 2004

The antibody-mix sandwich enzyme-linked immunosorbent assay (Ab-mix sELlS A) system was developed in order to simultaneously detect the extracellular polysaccharide (FPS) of Aspergillus, Penicillium, or Fusarium species using one detection system. The detection limit and detection range of the Ab-mix sELISA towards EPS of Penicilliun citrinum were not changed, and those towards Fusarium moniliforme EPS were changed a little compared to that of individual sandwich ELISA [9, 10]. The fungal culture filtrates of Aspergillus and Penicillium species showed nearly similar reactivity towards Ab-mix sELISA as that of sELISA using the MAb lB8 alone [9]. Also, the fungal culture filtrates of Fusarium species showed nearly the same reactivity towards Ab-mix sELISA as that of sELISA using the MAb lB8 alone [10]. Thus, this ELISA system showed that the three genera of molds, Aspergillus, Penicillium, or Fusarium, which are three major important molds producing mycotoxins in food or agricultural commodities, could be detected at the same time, using one detection system.

• 한국동물위생학회지
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• 제30권3호
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• pp.385-391
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• 2007

A novel enzyme linked immunosorbent assay (ELISA) is described and compared with other established serologic tests for bovine brucellosis, namely the rose bengal test (RBT), the complement fixation test (CFT), and the tube agglutination test (TAT) approved and used in Korea. A total of 109 bovine serum samples were tested using all the 4 assays and analyzed as to specificity, sensitivity, reproducibility and predictive value. The ELISA showed 100% agreement with the CFT. The least agreement between ELISA was observed with the TAT. The agreement between the ELISA and RBT was not significantly different from that observed between the CFT and the ELISA. It is concluded that the new assay would be a good candidate for routine serologic survey for brucellosis in Korea. A protocol combining the ELISA and the CFT would increase the power for detection of serologically positive individuals and herds.

• 센서학회지
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• 제30권5호
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• pp.337-341
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• 2021

In this paper, we describe the fabrication of a paper-based enzyme-linked immunosorbent assay (ELISA) to detect polygalacturonase (PG), which is used as a biomarker to determine whether a plant is infected with a disease. The proposed paper-based ELISA can analyze the concentration of PG in a short time using a small sample compared to the traditional ELISA, which is generally performed using a well plate. To increase the resolution of the sensor, we optimized the dilution ratio of the HRP-conjugated goat anti-rabbit IgG antibody and the dilution ratio of the anti-PG and HRP-conjugated goat anti-rabbit IgG antibodies. Furthermore, for quantitative analysis of PG concentration, Delta RGB analysis was conducted to detect color changes in the sensing window displayed by the PG samples at various concentrations. Based on the experiment, the fabricated paper-based ELISA could measure at least 0.25 ㎍ of PG and the measurement range was 0.25-2 ㎍. Therefore, the paper-based ELISA for detecting PG is expected to be able to determine the presence or absence of disease in crops at the infection stage in the future.

• 한국동물위생학회지
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• 제42권1호
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• pp.9-15
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• 2019

Transmissible gastroenteritis (TGE) is a disease confined to pigs of all ages, and can be a significant cause of economic loss in breeding herds, primarily because of the very high piglet mortality. The causative agent is a coronavirus, an enveloped positive strand RNA virus and closely related but non-enteropathogenic porcine respiratory coronavirus (PRCV). Although the TGEV has declined with its innocent relative, PRCV, further genome changes could not be excluded. Therefore, the herd-level immunity against this virus is important for the prevention of disease and should be carefully monitored. The aim of this study is to develop monoclonal antibody capture enzyme-linked immunosorbent assay (MAC-ELISA) which can rapidly and accurately determine a large numbers of serum samples for surveillance purpose, and to compare the ELISA with a TGEV-specific serum neutralization test. The MAC-ELISA was sufficiently achieved, and the comparison with the virus-specific serum neutralization assays for 713 sera from pig farms showed a high correlation ($r^2=0.812$, P<0.001). The specificity and sensitivity of MAC-ELISA for the serum neutralization test 91.9% and 91.6%, respectively, which means that the antibody detected by the MAC-ELISA could be said to be protective antibodies. In conclusion, the developed MAC-ELISA would be very helpful in evaluating protective antibodies against TGEV.

• 대한수의학회지
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• 제43권2호
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• pp.263-270
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• 2003

The objective of the present study was to investigate the use of fluid released from muscle samples as an alternative to serum for ELISA to detect classical swine fever(CSF) virus antibodies in slaughter pigs. The optimal correspondence between serum 1:20 OD values and muscle fluid OD values was achieved at a muscle fluid dilution of 1:2. Significant correlation was found between serum and neck muscle ELISA ($r_s=0.880$, p<0.0001, ${\kappa}=0.82$; specificity of 97.0% and sensitivity 90.6%). The semimembranous muscle showed similar correlation in CSF ELISA($r_s=0.877$, p<0.0001, ${\kappa}=0.75$; specificity of 94.1% and sensitivity 89.1%). High correlation was obtained between serum and mesenteric lymph node in the CSF ELISA ($r_s=0.937$, p<0.0001, ${\kappa}=0.87$; specificity of 97.1% and sensitivity 93.0%). Measmement agreement between serum ELISA and muscle fluid ELISA was calculated and expressed as limits of agreement. The correspondence of ELISA of serum and muscle fluid indicated limits of agreement. Above 95% of all muscle fluid values were distributed within this limits of agreement. Among the samples used for ELISA for detecting CSFV antibodies, mesenteric lymph node had the most correlation and agreement with serum ELISA. F-test for comparison of variances showed no significant difference between the serum and muscle fluid. In conclusion, muscle fluid is a useful postmortem alternative to serum to detect CSFV antibodies.

• Parasites, Hosts and Diseases
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• 제21권2호
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• pp.265-269
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• 1983

간흡충증 환자 혈청에서 Thin layer immunoassay에 의하여 항체를 정량적으로 측정하여 감수성과 특이성을 알아보고, 효소표식면역 법(ELISA)에 의한 성적과 비교하였다. 냉동 건조한 간흡충 성충의 생리식염수 추출물을 항원으로 사용하였고, TIA는 Elwing등(1976)의 방법으로, 효소표식면 역법은 Veiler등(1974)의 방법에 의해 실시하였다. 1. 간홉충중 환자 60명의 혈청에서 TIA zone 크기의 평균은 4.14mm였고, 3mm 이상을 양성으로 판정하였을 때 감수성은 100%, 특이성은 61.1%였다. 2. 효소표식면역 법에 의하면 ELISA치 0.8이상을 양성으로 판정하였을 때 감수성은 88.3%, 특이성은 87.3%였다. 3. 혈청내 IgG 농도는 TIA zone 크기와 상관이 없었으나 좌소표식면역법에 의한 ELISA치와는 상관이 있어 그 계수는 0.69였다. 4. TIA zone 크기는 효소표식면역 법에 의한 ELISA치와 그 분포에 있어 상관이 없었다.

• Applied Biological Chemistry
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• 제36권1호
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• pp.7-10
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• 1993

ELISA법을 zearalenone 생성균주 검색에 응용하였다. 먼저 zearalenone에 대한 항체를 500배 희석한 후 microtiter well에 $125\;{\mu}l$씩 주입하여 $40^{\circ}C$에서 overnight시켜 coating하고, 시료용액과 enzyme을 $37^{\circ}C$에서 30분간 반응시켰다. 배양후 washing buffer로 6회 세척한 다음 plate에 2,2'-azino-di-3-ethyl-benzthiazoline sulfonic acid(ABTS) 용액을 $100\;{\mu}l$씩 첨가하여 15분간 발색시킨 다음 반응 정지액 $100\;{\mu}l$씩을 가해 반응을 정지시키고 ELISA Reader로서 흡광도(410 nm)를 측정하였다. 그 결과 zearalenone 생성이 확인된 19 균주 중 분리균 R-5, C-46 및 S-134가 50 ng/ml 이상의 zearalenone을 생성하였다.

• Toxicological Research
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• 제27권2호
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• pp.125-131
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• 2011

Through the present study, we produced a monoclonal antibody against aflatoxin B1 (AFB1) using AFB1-carboxymethoxylamine BSA conjugates. One clone showing high binding ability was selected and it was applied to develop a direct competitive ELISA system. The epitope densities of AFB1-CMO against BSA and KLH were about 1 : 6 and 1 : 545, respectively. The monoclonal antibody (mAb) from cloned hybridoma cell was the IgG1 subclass with ${\lambda}$-type light chains. The $IC_{50}s$ of the monoclonal antibody developed for AFB1, AFB2, AFG1 and AFG2 were 4.36, 7.22, 6.61 and 29.41 ng/ml, respectively, based on the AFB1-KLH coated ELISA system and 15.28, 26.62, 32.75 and 56.67 ng/ml, respectively, based on the mAb coated ELISA. Cross-relativities of mAb to AFB1 for AFB2, AFG1 and AFG2 were 60.47, 65.97 and 14.83% in the AFB1-KLH coated ELISA, and 59.41, 46.66 and 26.97% in the mAb coated ELISA, respectively. Quantitative calculations for AFB1 from the AFB1-Ab ELISA and AFB1-Ag ELISA ranged from 0.25 to 25 ng/ml ($R^2$ > 0.99) and from 1 to 100 ng/ml ($R^2$ > 0.99), respectively. The intra- and inter-assay precision CVs were < 10% in both ELISA assay, representing good reproducibility of developed assay. Recoveries ranged from 79.18 to 91.27%, CVs ranged from 3.21 to 7.97% after spiking AFB1 at concentrations ranging from 5 to 50 ng/ml and following by extraction with 70% methanol solution in the Ab-coated ELISA. In conclusion, we produced a group specific mAb against aflatoxins and developed two direct competitive ELISAs for the detection of AFB1 in feeds based on a monoclonal antibody developed.

• 대한수의학회지
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• 제45권4호
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• pp.569-574
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• 2005

Enzyme-linked immunosorbent assay (ELISA) for detection of antibodies to reticuloendotheliosis virus (REV) at single serum dilution was standardized. REV HI, one of the Korean field isolates, was inoculated into chicken embryo fibroblast (CEF) cells and was harvested from the culture fluids and cells after 10 to 12 days. Viruses were purified by centrifugation at the $107,000{\times}g$ for 12 hours on 20, 30, 45% (W/V) sucrose gradient. Virus specific fraction was collected and used as ELISA antigen. To standardize ELISA, the optimal concentration of coating antigen ($1{\mu}g/well$) and conjugate (1/1000) was determined by corrected OD (OD value of positive serum-OD value of negative serum) and P/N ratio (OD value of positive serum/OD value of negative serum). To calculate ELISA titer by measuring absorbance at 1/400 single serum dilution, serum titrations were carried out for various sample sera together with standard positive and negative sera. The observed titers of serum samples were plotted against sample/positive (s/p) ratios at 1/400 serum dilution. From the above data, the ELISA titers could be calculated by the equation of $log_{10}$ ELISA titer = 2.2763 ($log_{10}$ s/p) + 3.482 (r = 0.93). For evaluating the sensitivity, the standardized method were compared with conventional agar gel immunodiffusion (AGID) test method using serum samples collected from REV infected field chicken flocks. Fifty seven of 60 samples (95%) were positive for REV by ELISA, whereas only 11 (18.3%) samples were positive by AGID test. This results suggested that the ELISA tests developed in this study could be used for detection of antibodies to REV with high sensitivity.