• 한국식품위생안전성학회지
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• 제8권4호
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• pp.205-214
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• 1993

The monoclonal antibody to chloramphenicol(CAP) was produced to develop an enzyme-linked immunosorbent assay(ELISA) for residual CAP. An immunogen(CAP-BSA) was prepared by immunogen, antibody titer was measured by indirect ELISA. Spleen cells form the immunized mouse were fused with SP2/OAg14 myeloma cells. Among hybridomas selected in HAT media, 6 clones shown high antibody titer to CAP were subjected to cloning by limit dilution, and all of the monoclonal antibodies(MCA1, 2, 3, 4, 5, 7 and 9) produced by each clone were identified as IgG1 by ELISA isotyping analysis. Competitive ELISA condition was established by using the purified monoclonal antibody MCA1 as primary antibody and CAP-HSA conjugate as coating antigen. Standard curve of CAP(n=28) showed that the lowest detection limit of CAP is 20ng/ml level. The cross-reactivities of the 6 monoclonal antibodies showed that CAP sodium succinate. CAP base, P-nitrophenol, and p-nitrobenzyl alcohol were 89∼178, 0.050∼2.237, 0.056∼0.794 and 0.013∼7.939%, respectively. No cross-reactivities were observed with phenylalanine, tyrosine, glutamine, thiamphenicol, neomycin, streptomycin, gentamicin, sulfamethazine, sulfathiazole, chlortetracycline and p-aminobenzoic acid.

• 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.

• 한국식품과학회지
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• 제46권3호
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• pp.269-275
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• 2014

메밀로부터 분리한 메밀단백질(BWP)을 토끼에게 면역하여 생산한 특이항체를 이용하여 가공식품 중 메밀의 검출을 위한 간접경합 효소면역측정법(ciELISA)을 확립하였다. 이때, BWP의 검출범위는 $0.05-100{\mu}g/mL$이었다. 특이항체 교차반응 결과, BWP, 메밀가루, 통메밀에 대한 반응성은 각각 100, 17.9, 11.8%를 나타내었으며 그 외 곡물 14종에 대한 반응성은 거의 나타나지 않아 메밀에 대한 특이성이 매우 높았다. 메밀가루는 $60-90^{\circ}C$에서 반응성이 평균 83.0%를 나타내었으며 $100^{\circ}C$에서는 44.5%로 반응성이 감소하였다. 메밀가루에 대한 spike test에서 메밀 생면, 삶은 면, 묵, 메밀곡물가루에서 평균 분석회수율이 각각 99.1, 98.6, 81.1, 104%로 매우 높게 나타났다. ciELISA에 의하여 22점의 시판시료 중 메밀의 함유량을 조사한 결과, 메밀 함유 표시된 12점에서 평균 분석회수율은 31.6%로 나타났으나 시료 중 메밀의 정성적 검출은 100%로 매우 우수하였다. 본 연구에서 개발한 ciELISA는 가공식품에 함유된 메밀을 검출하는데 매우 효과적으로 활용할 수 있을 것이다.

• 한국식품위생안전성학회지
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• 제11권4호
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• pp.287-290
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• 1996

To control the maximum residue level (MRL) for sulfamethazine (SMZ) residues in pork tissue, a microbial inhibition method is a regulatory screening assay method in Korea. Microwell plate-based competitive enzyme immunoassay (ELISA) kit is avalable for routine screening of SMZ residues in pork tissue. One ELISA kit is evaluated. Phosphate buffer extracts of samples fortified with SMZ at 0, 1, 5, and 10 ng/g were used in a recovery test of the kit. Market pork samples were assayed by the kit. Recovery of sulfamethazine was 104% at 10 ng/g. Intraassay variations and interassay variations for the kit were 7.70% and 5.76%, respectively. Concentration causing 50% inhibition of color development compared with blanks was 16.4ng. The violative pork samples with over MRL (0.1 $\mu\textrm{g}$/g) was 4 of 32 cases (12.5%) by used ELISA kit. This result indicates a possibility of the ELISA kit for screening test of SMZ residues in pork tissue, and still needs a comfirmatory assay for mandatory purposes.

• 대한수의학회지
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• 제33권3호
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• pp.539-545
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• 1993

This study was carried out the determine the progesterone concentration for serum by enzyme-linked immunosorbent assay(ELISA) in bovine adult at estrous, pregnant, after patuation and male, female calves of 1 month old, respectively. The results obtained were summarized as follows ; 1. The assay has a sesitivity of $0.1ng/m{\ell}$. 2. Intra and inter-assay coefficient of variation were 4.5%, 6.1~9.4% when used for the determination of progesterone in samples of bovine serum. 3. The percentages of recovery for progesterone added were between 88.0 to 88.9%. 4. Progesterone concentration of adult bovine serum at estrus, pregnant and after 1 day of parturition were $0.37{\pm}0.16$, $7.1{\pm}1.0$, $0.13{\pm}0.02ng/m{\ell}$, respectively. 5. There was no differences in serum progesterone concentration of calves both male($0.16{\pm}0.03ng/m{\ell}$) and female($0.15{\pm}0.04ng/m{\ell}$) on 1 month old. From these results, progesterone determination by ELISA is very applicable to detect of early pregnancy diagnosis, factorial analysis of reproductive disorder, and also reproductive physiological functions such as veterinary therapeutic measures and monitoring of cyclicity.

• Reproductive and Developmental Biology
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• 제34권3호
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• pp.161-167
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• 2010

Early diagnoses of pregnancy for animal such as swine and bovine is extremely important to increase income of a farmhouse and for the management of farm. For the development of immunoasaay system of pregnancy in swine, we report a competitive heterologous enzyme linked immunosorbent assay (ELISA) for the direct measurement of oestrone sulfate (E1S) in diluted urine using anti-E1G (glucuronide) monoclonal antibody which cross react with ElS. The principle of assay was based on the typical solid-phase competitive ELISA methods using E1G-HRP (horseradish peroxidase) as a tracer and E1S for standard. The method had a reasonable sensitivity for the detection of E1S with 0.15 ng/ml as a detection limit. The intra-assay and inter-assay precisions were raging coefficient of from 8.50~9.67% and 8.50~9.87%, respectively, which were quite acceptable. In a field trial with a group 37 sows (18 non-pregnancy and 19 pregnancy sows) after day 29~30 post service, the concentration of E1S were determined to be below 30 ng/ml in all non-pregnancy group and over 48 ng/ml in pregnancy group except one sample. The method described here, heterologous ELISA for the measurement of E1S in urine is good enough for monitoring the early pregnancy test of swine.

• 대한수의학회지
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• 제57권1호
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• pp.31-36
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• 2017

Japanese encephalitis (JE) is an important zoonosis caused by the mosquito-transmitted JE virus (JEV), which is a causative agent of reproductive failure in pregnant sows. Detection of JEV antibodies in swine is performed by hemagglutination inhibition (HI), virus neutralization (VN), and the plaque reduction neutralization test (PRNT). The most stringent PRNT is the 90% endpoint PRNT ($PRNT_{90}$). These conventional assays are difficult to carry out in diagnostic laboratories with insufficient instruments or cell culture systems. An alternative assay that is easily conducted and time efficient is required. In this study, we improved the indirect enzyme-linked immunosorbent assay (I-ELISA) with clarified antigen for the detection of JEV antibodies. The I-ELISA results obtained from 175 swine serum samples were compared with HI, VN, and $PRNT_{90}$ results. The sensitivity of I-ELISA was 91.8%, 95.0%, and 94.7% compared with HI, VN, and $PRNT_{90}$ results, respectively. The specificity of I-ELISA was 92.2%, 94.7%, and 94.7% compared with HI, VN, and $PRNT_{90}$ results, respectively. Moreover, the I-ELISA results were significantly correlated with the HI (r = 0.93), VN (r = 0.95), and $PRNT_{90}$ (r = 0.92) results. These results suggest that the improved I-ELISA is useful for serosurveillance of JEV in swine.