• Asian-Australasian Journal of Animal Sciences
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• v.28 no.5
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• pp.691-696
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• 2015

The objective of this study was to evaluate the quality of five commercial enzyme linked immunosorbent assay (ELISA) kits (A, B, C, D, and E) from different suppliers for detecting aflatoxin $B_1$ ($AFB_1$). $AFB_1$-free corn samples supplemented with different levels of $AFB_1$ (5, 10, and $20{\mu}g/kg$) were used as positive controls and 6 replicates of each control sample were tested to evaluate the accuracy and precision of these kits. In addition, we also evaluated the performance of these ELISA kits for $AFB_1$ in 30 feed samples, including corn, distillers dried grains with soluble, wheat samples, soybean meal, and poultry feed, which were verified by high performance liquid chromatography. Results showed that the coefficients of variation ranged from 1.18% to 16.22% in intra-plate and 2.85% to 18.04% in inter-plate for the determination of $AFB_1$. The half maximal inhibitory concentration for five kits ranged from 3.72 to $7.22{\mu}g/kg$. The quantitation limits of $AFB_1$ were all under the legal limit in China but somewhat inconsistent with kit instructions. Although the recovery rate of four of the five kits were either less than 90% or more than 110%, all these values were acceptable in practice. Two kits had high false positive rates (C and E). In conclusion, our results revealed that the qualities of five tested ELISA kits were significantly different.

• Environmental Analysis Health and Toxicology
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• v.5 no.3_4
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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.

• Korean Journal of Veterinary Research
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• v.37 no.3
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• pp.613-618
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• 1997

Direct competitive enzyme linked immunosorbent assay(ELISA) with monoclonal antibodies have been studied for quantitative determination of T-2 toxin from the mold corn. The T-2HS, T-2HS-BSA, T-2HS-HRP and monoclonal antibodies against T-2 toxin produced in the studies were qualified for quantitative ELISA test of T-2 toxin. The mean recovery rate from ground com spiked T-2 toxin was 83%. The meaning range of the T-2 test was 60ng to $2{\mu}g$. According to the recovery results with the com spiked T-2, the tests proved to be suitable in the screening of the moldy feed samples for the presence of T-2 toxin and will be able to become the basis of the ELISA test for the quantitative screening kits of T-2 toxin.

• Korean Journal of Food Science and Technology
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• v.32 no.3
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• pp.538-543
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• 2000

This study was conducted to develop an enzyme-linked immunosorbent assay(ELISA) for the determination of cooked goat meat. Muscle proteins were extracted from goat meat by heating at $98^{\circ}C$ for 15 min. Major thermostable(TS) protein, whose size and pI are 36 and 38 kDa and 4.5 respectively, were purified by DEAE-Sephadex A-50 and Sephadex G-75 column chromatography. The TS protein was immunized into rabbits in order to produce goat specific antibodies. Competitive indirect ELISA(ciELISA) was established by using the anti-TS antibody. The antibody showed high reactivity toward the TS antigen and the boiled goat meat extract but it did not show any reactivities toward extracts of boiled chicken, pork, lamb, and beef. Thus, this ciELISA developed in this study could be applicable to identify goat species from cooked meat.

• The Journal of the Korean Society for Microbiology
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• v.22 no.2
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• pp.117-123
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• 1987

Previous studies have been performed for the sero-identification of selected species of Bacteroides by immunofluorescence antibody techniques and enzyme-linked immunosorbent assay using species-specific rabbit antisera to B. gingivalis, B. intermedius, and B. melaninogenicus. However, these studies have not commented on the serological cross-reactivity between these 3 species of black- pigmented Bacteroides. For the cross-reactivity study, antisera to B. gingivalis ATCC 33277, B. intermedius ATC C25261 and B. asaccharolyticus ATCC 25260 were raised from rabbits. Preliminary study for observing the cross-reactivity between these species was performed by indirect immunoflourescence technique. Immunoabsorption of the antisera was done with bacterial cells from the other species and the species-specificity of the antisera was conformed by the absence of reactivity with bacterial strains from the other species by indirect immunofluorescence technique and enzyme-linked immunosorbent assay. Three representative unabsorbed antisera cross-reacted strongly with cells from the other species. Especially, anti-B. asaccharolyticus ATCC 25260 antiserum showed a strong cross-reactivity with B. gingivalis ATCC33277. After immunoabsorption of 3 representative antisera with the other species, the cross-reactivity was found only between B. gingivalis ATCC 33277 and B. asaccharolyticus ATCC 25260. Further study would be necessary to clarify the cross-reactivity between important oral black-pigmented Bacteroides from subgingival plaque or bacterial colonies for rapid identification.

• Animal Bioscience
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• v.34 no.12
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• pp.1995-2002
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• 2021

Objective: Detection of adulteration in processed meats is an important issue for some countries due to substitution of beef with a cheaper source of protein like poultry. In this study, the presence of chicken meat was investigated using real-time polymerase chain reaction (real-time PCR) and enzyme-linked immunosorbent assay (ELISA) techniques to verify adulteration of fermented sausage samples. Methods: A total of 60 commercial samples were collected from 20 establishments in three replicates including 10 fermented sausage manufacturers and 10 butchers to investigate the presence of chicken meat with the sequential use of real-time PCR and ELISA techniques. In addition, pH, moisture content, water activity and color values of the samples were determined. Results: Both real-time PCR and ELISA showed agreement on the presence or absence of chicken meat in 55 out of 60 fermented sausage samples and chicken meat was identified with both methods in 16 samples. Five samples produced inconsistent results for the presence of chicken meat in the first run. Nevertheless, the presence of chicken meat was verified with both methods when these samples were analyzed for the second time. In addition, the average physico-chemical values of the fermented sausage samples tested positive for chicken meat were not significantly different from some of those fermented sausage samples tested negative for the chicken meat. Conclusion: The sequential use of real-time PCR and ELISA techniques in fermented sausages could be beneficial for the government testing programs to eliminate false negatives for detection of adulteration with chicken meat. Furthermore, consumers should not rely on some of the quality cues including color to predict the adulteration of fermented sausages with chicken meat since there were no statistical differences among some of the samples tested positive and negative for chicken meat.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.43 no.4
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• pp.228-233
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• 1998

Seed storage proteins have been used for studying biochemical genetics and end-use quality aspects. We conducted enzyme-linked immunosorbent assay (ELISA) and one-dimensional SDS-PAGE (1D SDS-PAGE) to evaluate different cereal crop species and Korean wheat lines for rye secalin proteins. The antisecalin antibody showed consistent specificity for rye secalin with little cross-reactivity to gliadins. Immunological cross-reactivities measured by the ELISA technique using competition assay showed significant differences of absorbance among rye, triticale, wheat-rye translocated wheat and non-translocated wheat. The absorbance values were lowest in rye followed by triticale, translocated wheat and non-translocated wheat. The ELISA for discrimination of wheat-rye translocation on the basis of antigen-antibody reactivity showed that none of the Korean wheat lines possessed 1RS and secalin proteins. The competitive ELISA experiment demonstrated specific determination for secalin that was originated from rye chromosomal parts. The result of 1D SDS-PAGE for identifying rye secalin subunits showed all three rye specific secalin protein subunits (75 KDa, 45 KDa, and 40 KDa) for rye and triticale, and 1RS specific secalins (45 KDa and 40 KDa) for 1AL/1RS and 1BL/1RS translocated wheats. All Korean wheats were lacking 1RS of rye chromosome and secalin.

• Clinical and Experimental Reproductive Medicine
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• v.40 no.1
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• pp.12-22
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• 2013

Objective: We investigated the norepinephrine transporter (NET) expression in normal and pre-eclamptic placentas and analyzed the invasion activity of trophoblastic cells based on norepinephrine (NE)-NET regulation. Methods: NET and NE expression levels were examined by western blot and enzyme-linked immunosorbent assay, respectively. Trophoblast invasion activity, depending on NE-NET regulation, was determined by NET-small interfering RNA (siRNA) and NET transfection into the human extravillous trophoblast cells with or without NE treatment and invasion rates were analyzed by zymography and an invasion assay. Results: NET mRNA was expressed at a low level in pre-eclamptic placentas compared with normal placentas and NE concentration in maternal plasma increased significantly in pre-eclamptic women compared to normal pregnant women (p<0.05). NET gene upregulation and NE treatment stimulated trophoblast cell invasion up to 2.5-fold (p<0.05) by stimulating matrix metalloproteinase-9 activity via the phosphoinositol-3-kinase/AKT signaling pathway, whereas NET-siRNA with NE treatment reduced invasion rates. Conclusion: NET expression is reduced by inadequate regulation of NE levels during placental development. This suggests that a complementary balance between NET and NE regulates trophoblast cell invasion activities during placental development.

• Korean Journal of Veterinary Research
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• v.31 no.4
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• pp.403-409
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• 1991

This experiment was carried out to develop a sensitive, rapid, solid-phase microtitre plate assay of progesterone using the monoclonal antibody to this hormone. Monoclonal antibody to progesterone was much higher titre and binding affinity about 10 times than conventional polyclonal antibody to progesterone. Dot-blot analysis of monoclonal antibody revealed a single precipitation band when reacted with anti-mouse IgM and anti-mouse K. A competitive reaction was used with a reaction time of 2 hours. The standard dose-response curve was linear through 1,000pg/well. This ELISA system approach is applicable to evaluation for the rapid assessment of luteal function and reproductive status in both clinical and research in a wide variety of species.

• Ocean and Polar Research
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• v.25 no.3
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• pp.249-256
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• 2003

An immunological method was developed in this study to quantify reproductive output of the female butter clam, Saxidomus purpuratus. A clam egg-specific polyclonal antibody was developed using the purified butter clam egg as an antigen. An indirect Enzyme-Linked Immunosorbent Assay (ELISA) was used in quantitative measurement of the eggs. Size of the butter clam eggs ranged from $70.81{\pm}7.52{\mu}m$ in histology or $88.56{\pm}11.31{\mu}m$ in intact eggs. The predominant egg constituent was protein (37.44%), followed by lipids (11.40%) and carbohydrates (9.68%). The SDS-PAGE showed that the egg proteins are composed of several peptides of molecular weights consisting of 247, 200, 99, 91, 54 and 47 kDa. ELISA indicated that the clams collected from Geoje Island in May 2002 produced 8.2 to 26.8% of their body weight as eggs or 9,307,309 to 31,156,333 with a mean of 16,931,893 eggs per individual clam. The results of this study thus suggest that indirect ELISA using rabbit anti-clam egg IgG as a primary antibody is a rapid, affordable and sensitive method to assess reproductive output of 5. purpuratus and possibly other bivalves using a small amount of eggs.