• Title/Summary/Keyword: ELISA

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A survey on diseases to improve productivity in 1-day-old chicks of broiler farms (농가 생산성 증대를 위한 육계 초생추 질병 실태조사)

  • Koh, Won-Seuk;Um, Sung-Shim;Cho, Bum-Jun;Kim, A-Rum;Lee, Byong-Jong;Lee, Seong-Hyo;Bae, Joung-Jun
    • Korean Journal of Veterinary Service
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    • v.30 no.3
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    • pp.329-338
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    • 2007
  • Samples collected from 15 broiler farms(47 flocks, 920 1-day-old chicks) during March to December, 2006, To survey serum antibody titers of NDV, IBDV and MG/MS, the antibodies of ND viruses were detected by HI test and ELISA, against antibodies of IBD viruses and MG/MS by ELISA. The antibody titers of NDV showed 6.4, HI and 6,968, ELISA, respectively. The rate to below protective antibody levels(${\ge}5$, HI and ${\ge}1,000$, ELISA) were 8%, HI, 5%, ELISA, specially, Baeksemi were 22%, HI, 14%, ELISA. The rate of positive by ELISA showed 99%(914/920). The ELISA titer of IBDV showed mean titer 3,890. The rate of positive were 93% (857/920), specially, Baeksemi were 84%. The ELISA titers of MG/MS showed mean titer 5,666. The rate of positive were 78% (715/920) and 100%, Abor-Acre, 97%, Baeksemi, respectively. The antibodies not detected from 18%, ELISA titers was varied from 500 to 20,000. At antimicrobial susceptibility of E coli, Staphylococcus spp and Salmonella spp isolated from 1-day-old chicks, E coli were susceptible to AmC, AM, NOR, SXT, ENR, CIP, Staphylococcus spp were susceptible to AmC, SXT, AM, ENR and Salmonella spp were susceptible to AM, AmC, SXT and P.

Detection of Xanthomonas axonopodis pv. citri on Citrus Fruits Using Enzyme-Linked Immunosorbent Assay

  • Jin, Kyoung-Sik;Kang, Ik-Beom;Ko, Kyoung-Il;Lee, Eun-Seob;Heo, Jong-Young;Kang, Young-Kil;Kim, Byung-Ki
    • The Plant Pathology Journal
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    • v.17 no.1
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    • pp.62-66
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    • 2001
  • Detection of Xanthomonas axonopodis pv. citri (Xac) on citrus fruits for exporting is usually made by bacteriophage test (BPT) to demonstrate the pathogen-free status. BPT has rather time-consuming and complicate procedures for dealing with massive samples to be inspected. In this study, enzyme-linked immunosorbent assay (ELISA) was applied to detect Xac on fruits, and compared with BPT. In ELISA, positive reactions occurred in the bacterial densities of $3\times10^5$ cells/ml or more. To detect the bacterial infection on citrus fruits with a density of lower than $3\times10^5$ cells/ml, the bacterial suspensions were mixed with fruit rinse water and incubated in broth medium. Ordinary peptone sucrose broth (PSB) was not a proper medium for increasing Xac density specifically enough to be detect by ELISA. On the other hand, modified PSB (MPSP) amended with Fe-EDTA (0.25 g/$\ell$) and 2.5% potato-dextrose broth sufficed to differentiate uninfected and infected citrus fruits by ELISA after 24 h incubation of the fruit rinse water. Using various citrus samples from infected and uninfected fields, efficiencies in detecting Xac on fruits were compared between ELISA and BPT. For infected fruits samples, ELISA detected Xac by 100%, while BPT by about 44%, indicating that the detection efficiency was improved by 23.5% by ELISA, compared to BPT. In addition, ELISA has simpler procedures for testing and is less time-consuming than BPT, suggesting that ELISA may be accurate and simple method to detect Xac on citrus fruits.

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Establishment and application of a solid-phase blocking ELISA method for detection of antibodies against classical swine fever virus

  • Cao, Yuying;Yuan, Li;Yang, Shunli;Shang, Youjun;Yang, Bin;Jing, Zhizhong;Guo, Huichen;Yin, Shuanghui
    • Journal of Veterinary Science
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    • v.23 no.5
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    • pp.32.1-32.11
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    • 2022
  • Background: Classical swine fever (CSF) is a severe infectious disease of pigs that causes significant economic losses to the swine industry. Objectives: This study developed a solid-phase blocking enzyme-linked immunosorbent assay (spbELISA) method for the specific detection of antibodies against the CSF virus (CSFV) in porcine serum samples. Methods: A spbELISA method was developed based on the recombinant E2 expressed in Escherichia coli. The specificity of this established spbELISA method was evaluated using reference serum samples positive for antibodies against other common infectious diseases. The stability and sensitivity were evaluated using an accelerated thermostability test. Results: The spbELISA successfully detected the antibody levels in swine vaccinated with the C-strain of CSFV. In addition, the detection ability of spbELISA for CSFV antibodies was compared with that of other commercial ELISA kits and validated using an indirect immunofluorescence assay. The results suggested that the spbELISA provides an alternative, stable, and rapid serological detection method suitable for the large-scale screening of CSFV serum antibodies. Conclusions: The spbELISA has practical applications in assessing the vaccination status of large pig herds.

Development of an Indirect Enzyme-Linked Immunosorbent assay for Rapid Detection of Adulteration of Food Allergen Mackerel in Processed Marine Foods (수산가공식품 중 알레르겐 고등어 혼입여부 신속 검출을 위한 간접효소면역분석법의 개발)

  • Lee, Jeong-Eun;Kim, Ah-Yoon;Kim, Sol-A;Kim, Hyo-In;Park, Ji-Hye;Shim, Won-Bo
    • Journal of Food Hygiene and Safety
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    • v.33 no.3
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    • pp.185-192
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    • 2018
  • The purpose of this study was to develop an indirect enzyme-linked immunosorbent assay (indirect ELISA) based on a monoclonal antibody (MAb) that is specific to mackerel thermal stable-soluble protein (TSSP), that can be used for the rapid detection of mackerel in processed marine foods. Among the four MAbs (3A5-1, 2, 9, and 12) developed in previous studies, the 3A5-2 MAb that showed high specificity and sensitivity were selected and used to develop the indirect ELISA method. The detection range of the indirect ELISA was 0.02%-0.001% and the detection limit of 0.001% was shown. No cross-reaction to other marine products and food ingredients was observed by the indirect ELISA. Processed marine foods containing mackerel with ${\geq}0.3$ O.D. value at 405 nm were estimated as positive samples by the indirect ELISA. Therefore, the indirect ELISA can be used as a rapid and sensitive method to identify mackerel authenticity and adulteration in processed marine foods.

Development of an ELISA kit for the detection of residual sulfadimethoxine in edible animal products (축산물 잔류 sulfadimethoxine 검출용 ELISA kit 개발)

  • Kim, Woo-taek;Kim, Seong-hee;Yoon, Byoung-su;Lim, Yoon-kyu
    • Korean Journal of Veterinary Research
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    • v.40 no.3
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    • pp.601-609
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    • 2000
  • An enzyme linked immunosorbent assay (ELISA) was developed to screen residues of sulfadimethoxine (SDM) in edible animal products. An indirect competitive ELISA was allowed to compete with rabbit anti-SDM for binding to a limited amount of SDM-gelatin conjugate and SDM in serum samples. Sera was diluted 20 times with phosphate buffered saline (PBS) and boiled for 5 minutes to destruct immunoglobulins of serum. Detection limit of this competitive ELISA for SDM was 0.1 ppb or less. Among eight sulfonamide analogues tested for specifity, only sulfamonomethoxine showed significant cross-reaction in the assay. The EC-50 value for sulfamonomethoxine was 3.5 ppm. Recovery of SDM in spiked serum samples between 100 ppb and 500 ppb ranged from 110.7% to 128.9%.

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Quantitative Analysis of Phosphinothricin-N-acetyltransferase in Genetically Modified Herbicide Tolerant Pepper by an Enzyme-Linked Immunosorbent Assay

  • Shim, Youn-Young;Shin, Weon-Sun;Moon, Gi-Seong;Kim, Kyung-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.17 no.4
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    • pp.681-684
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    • 2007
  • An immunoassay method was developed to quantitatively detect phosphinothricin-N-acetyltransferase (PAT) encoded by the Bialaphos resistance (bar) gene in genetically modified (GM) pepper. The histidine-tagged PAT was overexpressed in Escherichia coli M15 (pQE3l-bar) and efficiently purified by $Ni^{2+}$ affinity chromatography. A developed sandwich enzyme-linked immunosorbent assay (S-ELISA) method (detection limit: $0.01{\mu}g/ml$) was 100-fold more sensitive than a competitive indirect ELISA (CI-ELISA) method or Western blot analysis in detecting the recombinant PAT. In real sample tests, PAT in genetically modified herbicide-tolerant (GMHT) peppers was successfully quantified [$4.9{\pm}0.4{\mu}g/g$ of sample (n=6)] by the S-ELISA method. The S-ELISA method developed here could be applied to other GMHT crops and vegetables producing PAT.

The Study on the Establishment of Specific ELISA for the Detection of Fish Metallothionein (어류 Metallothionein에 대한 특이적 면역 효소 측정법의 확립에 관한 연구)

  • 황갑수
    • Environmental Analysis Health and Toxicology
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    • v.11 no.1_2
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    • pp.11-17
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    • 1996
  • The purpose of this experiment is to estabilish the sensitive and specific ELISA (enzyme linked immunosorbent assay) system for the detection of fish metallothionein (MT). Silver carp were injected with CA of 1-8mg/kg body wt. 4 times during 10 days. Silver carp was very tolerant species to CA. Cd induced MT in liver was seperated and purified by gel filtration chromatography and ion exchange chromatography and identified by spectrophotometry, native gel electrophoresis and western blot analysis. The rabbit antiserum was produced by immunizing rabbit with lyophilized MT, and the competitive ELISA system was estabilished for the detection of fish MT. In the present ELISA system, the detection limit was about 33 ng/ml. When this ELISA system was employed to determine the MT level in the supernatant sample of fish liver homogenate, the reaction curve showed a good parallel corelationship with the calibration curve over a certain dilution range. The results indicate that the competitive ELISA can be a useful tool for the detection of fish MT in the toxicological study and the evaluation of water pollution.

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Comparative Analysis of Screening Results from Various ELISA Formats Used for Detection of Anti-Erythropoietin Antibodies in Korean Patients

  • Ha, Sung-Kyu;Yang, Seung-Ju;Shin, Sug-Kyun;Jo, Young-Il;Baek, Kyung-Min;Hong, Seung-Hwa;Pack, Seung-Pil;Kim, Sung-Jo;Heo, Tae-Hwe
    • Biomolecules & Therapeutics
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    • v.18 no.2
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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.

Microplate Enzyme-Linked Immunosorbent Assay for Bovine Virus Antibody (우백혈병(牛白血病) Virus 항체측정(抗体測定)을 위한 효소면역법(酵素免疫法))

  • Choi, Won Pil
    • Current Research on Agriculture and Life Sciences
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    • v.1
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    • pp.195-199
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    • 1983
  • A microplate enzyme-linked immunosorbent assay (ELISA) for detecting antibodies to bovine leukemia virus(BLV) is described and compared its sensitivity with that of the agar gel immunodiffusion test (ID) with BLV glycoprotein (gp) antigen using 263 sera collected in Korea and Japan. There was 98.5 per cent agreement between ELISA and ID when ELISA value, the value of tested serum(T) was devided with that of standard negative seurm(N) after the value of control was eliminated from T and N (T-C/N-C), of 1.5 or greater was considered positive. One hundred and forty four (99.6%) of 145 sera which were positive by ID were greater than 1.5 by ELISA, and 115 (97.5%) of 118 sera which were negative by ID were less than 1.5 by ELISA. As a result, it suggest that the ELISA test using BLV-gp antigen provides a useful serological tool for the diagnosis of BLV infection.

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Development of Enzyme-Linked Immunosorbent Assay for Rapid and Sensitive Analysis of Biotin (Biotin의 분석을 위한 효소면역측정법(ELISA)의 개발)

  • 이경애;손동화;고영태
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.27 no.6
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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.

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