• Title, Summary, Keyword: ELISA

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Comparison of ELISA and HI titers in broiler chicks vaccinated with infectious bronchitis virus and Newcastle disease virus (전염성기관지염 및 뉴캣슬병 백신을 접종한육계에서 ELISA 및 HI 항체가 비교)

  • 고원석;이정원;곽길한;권정택;송희종
    • Korean Journal of Veterinary Service
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    • v.24 no.1
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    • pp.21-29
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    • 2001
  • To compare of serum antibody titers using ELISA and HI, serum samples were collected from 100 breeders and their progeny 550 broilers. The breeders and broilers were vaccinated with infectious bronchitis(IB)- and Newcastle disease(ND)-viruses according to general vaccination program. The antibodies in serum samples against IB and ND viruses were detected by enzyme-linked immunosorbent assay(ELISA) using commercial ELISA kit and hemagglutination inhibition(HI) test. Geometric mean titer(GMT) of ELISA and In titers were monitored from 1-day-old to 35-day-old broilers and compared to those of breeder chickens. The antibody titers of breeders vaccinated with ]B virus showed 47,800, ELISA and 7.2, HI, respectively. Progeny chicks, 1-day-old, vaccinated with IBV showed high antibody titers than those of breed chickens. Those chicks were maintained protective antibody levels until 11-day-old. From 14-day-old, the antibody level decreased below protective levels. In ND, breeders serum antibody titers ELISA and Eiu were 30,200 GMT and 8.7 HI titer, respectively. On 1-day-old chicks, antibody levels was decreased to half in ELISA(16,270) compared with those of breeders, but In titers was 7.4. Progeny broilers, protective antibody level was maintained until 14- day-old by ELISA, but at 11-day-old by HI titers. After then, ND antibody titer was continuously decreased underdefense level. These result indicated that the ELISA method be more sensitive than HI titration to detect serum antibody level for IBV and NDV.

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Efficacy of ELISA for measurement of protective newcastle disease antibody level in broilers (육계의 뉴켓슬병 방어역가 측정에 있어서 ELISA 검사법의 효용성)

  • Kim, Jong-Nyeo;Heo, won;Mo, In-Pil
    • Korean Journal of Veterinary Research
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    • v.46 no.3
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    • pp.185-196
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    • 2006
  • Newcastle disease (ND) is a highly contagious disease of poultry that can cause severe economic losses throughout the world. Vaccination has been used for a long time and proved as one of the most effective method to reduce the economic loss due to ND virus infection, The measurement of antibody titer such as hamagglutination-inhibition (Hl) test with sera has been used as a useful method to evaluate the immunity leve of host. However, Hl test is gradually being replaced by the enzyme linked-immunosorbent assay (ELISA), To evaluate the efficacy of ELISA in the chickens vaccinated with different procedure, present study has been performed. After SPF chicks and commercial broilers were vaccinated with different kinds of live vaccines such as V4, VG/GA and/or Bl at various time, the antibody level has been measured using both HI test and ELISA. Challenge test with velogenic viscerotropic NDV was also performed to measure the protective level of antibody. In the SPF chickens, the mean ELISA titer after vaccination and survival rate after challenge was increased and correlated with days post inoculation. More than 80% of chickens with higher than 1,000 ELISA titer after vaccination were survived after challenge with velogenic ND virus and had good correlation between survival rate and antibody titier. In commercial broiler chickens, most of them at market age had low level of ELISA titer regardless of the number of vaccination, and had a low correlation between survival rate and ELISA titer. However, the ELISA titer of remaining birds after challenge was increased. This result indicated that ELISA titer had good response against velogenic NOV infection compared to Hl titer.

Indirect ELISA Method for Measurement of Lactoperoxidase using IgY Antibody (IgY 항체를 이용하여 Lactoperoxidase 정량을 측정하기 위한 Indirect ELISA 방법의 개발)

  • 이승배;최석호;최재원
    • Food Science of Animal Resources
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    • v.24 no.2
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    • pp.182-188
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    • 2004
  • To determine the concentration of Lactoperoxidase (LPO), an indirect enzyme-linked immunosorbant assay(ELISA) was developed. Anti-LPO egg yolk immunoglobulin(IgY) was transferred to egg yolk by immunizing of Brown hens with LPO. The titer of purified anti-LPO IgY was 1: 520,000. The immunological response of anti- LPO IgY with ${\alpha}$-lactalbumin, ${\beta}$-lactoglobulin, casein and lysozyme were evaluated, resulting that the anti-LPO IgY found to be a specific antibody toward LPO and no cross-reaction was observed against ${\alpha}$-lactalbumin, ${\beta}$-lactoglobulin, casein, and lysozyme in double immunodiffusion test and ELISA test. In indirect ELISA method, coating concentration of LPO and dilution rate of anti-LPO IgY was 0.25$\mu\textrm{g}$/mL and 1:8,000 respectively. Sensitivity in the standard curve of LPO was ranged from 0.01 to 1$\mu\textrm{g}$/mL using anti-LPO IgY.

Excretory-secretory antigen is better than crude antigen for the serodiagnosis of clonorchiasis by ELISA

  • Choi, Min-Ho;Park, Il-Chan;Li, Shun-Yu;Hong, Sung-Tae
    • The Korean Journal of Parasitology
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    • v.41 no.1
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    • pp.35-39
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    • 2003
  • Although stool examination is the standard diagnostic method of clonorchiasis, serodiagnosis by ELISA using crude antigen is now widely used because of its convenience. However, ELISA diagnosis still suffers from cross-reactions, and therefore there is a need to improve the present conventional ELISA. The present study was undertaken to evaluate the diagnostic value of ELISA using excretory-secretory antigen (ESA) instead of crude antigen (CA) of Clonorchis sinensis. The diagnostic sensitivity of ELISA using excretory-secretory antigen was 92.5%, which was higher than that of ELISA using crude Clonorchis sinensis antigen (88.2%). In addition, the specificity of excretory-secretory antigen was found 93.1% while that of crude antigen was 87.8%. In summary, Clonorchis sinensis ESA was found to be a better serodiagnostic antigen than CA for ELISA.

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.

Intraerythrocytic culture and development of serological diagnostic tests of Babesia gibsoni 1. Indirect fluorescent antibody test and enzyme-linked immunosorbent assay for antibody detection of Babesia gibsoni infections in dogs (Babesia gibsoni의 적혈구내 배양법과 진단법 개발에 관한 연구 1. Babesia gibsoni 진단을 위한 간접형광항체법(IFAT)과 효소표지면역검사법(ELISA))

  • Suh, Myung-deuk;Shin, Yong-seung
    • Korean Journal of Veterinary Research
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    • v.37 no.3
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    • pp.583-593
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    • 1997
  • Indirect fluorescent antibody test(IFAT) and enzyme-linked imuunosorbent assay (IgG-ELISA) as serological diagnostic tools were conducted to evaluate the usefulness for diagnosis of canine babesiosis infected with Babesia gibsoni in domestic various dog breeds, american pit bullterrier, military shepherd, and mongrel dogs. The results obtained from this study were abstracted as follows. The nonionic detergent Triton X-100 and absorbent bio-bead $SM_2$ were useful reagents for the preparation of pure merozoite antigen of B gibsoni to be used in ELISA. The optimum reaction in ELISA was shown when the protein concentration of ELISA antigen was measured as 625ng/ml and the conjugate concentration was diluted into 1/6000 fold. The average OD value of ELISA in sera determined with negative responses in IFAT was measured as $0.255{\pm}0.051$(490nm) and the cut - off value of OD was determined as 0.399(490nm). The serum antibodies in both of IFAT and ELISA were detected on one week after artificially infected with B gibsoni and these high antibody titers, 512X in IFAT and 1024X in ELISA, were long lasted until 15 weeks after infection. The reproducibility of reaction and stability of the antigen absorbed microtitration polystyrene plate preserved in $4^{\circ}C$ refrigerator and $-20^{\circ}C$ freezer, respectively could be lasted until 135 days after storage. The positive rates in IFAT by dog breeds were shown 8.1%(60/744 heads) in mongrel dogs, 81.3%(78/96 heads) in american pit bullterrier and 15.6%(15/96 heads) in military shepherd, while the positive rate in ELISA shown 17.6%(131/744 heads) in mongrel dogs, 83.3%(80/96 heads) in american pit bullterrier and 36.5%(35/96 heads) in military shepherd, respiectively. In the total of 936 heads surveyed with IFAT and ELISA the positive rates in IFAT and ELISA were 16.4%(153/936 heads) and 26.3%(246/936 heads), respectivily. Agreement of reactions between IFAT and ELISA was shown 82.4% in 936 dog sera. The specificity and sensitivity of ELISA reaction were 83.5% and 76.5%, respectively. From the conclusion obtained in this study it was evaluated that IFAT and ELISA were useful as highly specific, sensitive and stable serelogical tools for the diagnosis of canine babesiosis in Korea.

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Enzyme-Linked Immunosorbent Assays of Pseudomonas tolaasii, a Bacterial Brown Blotch Pathogen of Oyster Mushroom. (느타리버섯 세균성갈반병균 Pseudomonas tolaasii의 효소면역검출법)

  • 이향범;전낙범;손동화;유승헌
    • Microbiology and Biotechnology Letters
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    • v.26 no.3
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    • pp.238-243
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    • 1998
  • For simple and rapid detection of Pseudomonas tolaasii (PT), a bacterial brown blotch pathogen of oyster mushroom, enzyme-linked immunosorbent assays (ELISA) were developed. To produce specific antibody, PT ($5{\times}10^7$ cfu) and Freund's adjuvant were subcutaneously immunized into rabbits several times. By using the antiserum showing the highest titer, we established noncompetitive and competitive ELISA's. Standard curves of the ELISA's showed that the detection limits were $2{\times}10^2$cfu/ml and $3{\times}10^2$cfu/ml, respectively When investigated by noncompetitive ELISA, cross reactivities of the anti-PT antibodies against P. agarici, P. reactans, and other fluorescent Pseudomonas spp. were very low (<1/10$^3$), but those against P. solanacearum, Erwinia chrysanthemi, Streptococcus mutans, Xanthomonas citri, and a fungus Fusarium oxysporum were almost none. However, when investigated by competitive ELISA, the reactivities against any other strains except PT were almost none. When the ELISA's were applied to 18 strains derived from mushrooms in order to identify PT, only 11 strains showing both pathogenicity and white line reactivity were obviously positive. These results showed that the ELISA's could be convenient tools to detect PT in accordance with existing methods.

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A Comparative Study of Serologic Methods for Detection of Mumps Antibody in Korean Children (한국 소아의 Mumps 바이러스 항체보유에 관한 혈청학적 진단방법의 비교)

  • Park, Hae-Kyung
    • The Journal of the Korean Society for Microbiology
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    • v.21 no.4
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    • pp.473-480
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    • 1986
  • Mumps is an extremly common infectious disease affecting predominantly young children hut it is not a severe disease in terms of mortality. One hundred and two sera from infants of 3 different groups which are vaccinated, unvaccinated and unknown were detected to mumps antibody. The tests used were Complement Fixation(CF) test, Single Radial Hemolysis(SRH) test, Hemagglutination Inhibition(HI) test, Enzyme Linked Immunosorbent Immunoglobulin G(ELISA IgG) test, Enzyme Linked Immunosorbent Immunoglobulin M(ELISA IgM) test. 1. The rate of positivity for mumps antibody in 102 sera wera 89.16%(74/83) by Hl test, 68.83%(53/77) by ELISA IgG test, 64.58%(62/96) by SRH test, 63.24%(43/68) by ELISA IgM test and 50.00%(49/98) by CF test. 2. The rate of positivity by 5 tests for 55 sera turned out to be very similar with above results respectively. 3. The correlation coefficients(r) between ELISA IgG test ant H1 test, ELISA IgG test and ELISA IgM test were 0.34(P<0.0l) and 0.31(P<0.02), respectively. 4. The percentage of apparently natural infection of mumps seemed to be 65.15%(43/66) in infants. 5. Seroconversion rate of mumps by vaccination were 90.91%(10/11). 6. Among the 53 infants who were tested with ELISA IgG 15 were below 15 months age of(28.30%) and this percentage may be taken as a suggestion that mumps vaccination should be given earlier than present practice. 7. ELISA IgG test was found very sensitive and recommendable method for large scale screening for the presence of antibody to mumps.

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Enzyme-Linked Immunosorbent Assay for Detection of Nivalenol (Nivaleno의 검출을 위한 효소 면역 측정법)

  • 손동화;이향범;곽보연;김수호;권창희
    • Journal of Food Hygiene and Safety
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    • v.13 no.2
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    • pp.129-134
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    • 1998
  • To develop an enzyme-linked immunosorbent assay (ELISA) for nivalenol (NIV), we produced polyclonal antibodies against tetraacetyl nivalenol (Ac4-NIV) and established ELISA conditions. Ac4-NIV-hemisuccinate conjugated to bovine serum albumin (Ac4-NIV-HS-BSA) was immunized with Freund's adjuvants into rabbits subcutaneously several times. By use of the antiserum showing the highest titer and Ac4-NIV-HS-HRP conjugate, we established competitive direct ELISA (cdELISA). Standard curve of cdELISA showed that the detection range of Ac4-NIV was about 10~5,000 ng/ml (ppb). The cross-reactivities of the polyclonal antibody towards Ac4-NIV and acetyl T-2 were 100 and 70% respectively, and those towards NIV, deoxynivalenol, 3-acetyl deoxynivalenol, 15-acetyl deoxynivalenol, triacetyl deoxynivalenol, fusarenonX, and T-2 were less than 0.1%. When cdELISA was applied to NIV-spiked corns followed by extraction with 70% acetonitrile and acetylation with acetic anhydride in pyridine, the recovery rates of the Ac4-NIV were 108, 143, and 70% (average, 107%) in the levels of 100, 300, and 1,000 ng/g (ppb), respectively.

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Detection of Salmonella in Milk by Sandwich ELISA using Anti-Outer Membrane Protein Immunoglobulins (Anti-Outer Membrane Protein 면역단백질을 이용한 Sandwich ELISA 방법에 의한 우유 내 Salmonella의 검출)

  • 최석호
    • Food Science of Animal Resources
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    • v.24 no.2
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    • pp.176-181
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    • 2004
  • The specificity of sandwich enzyme-linked immunosorbent assay (ELISA) to detect Salmonella in milk was determined in this study. The antibodies used in sandwich ELISA were egg yolk immunoglobulin G (IgY) obtained after immunization of hen with outer membrane protein (OMP) fraction from Salmonella typhimurium and rabbit IgG obtained after immunization of rabbit with the purified OMP with the molecular weight of 40,000. The immunoblot assay showed that the IgY reacted strongly with OMP with the molecular weight of 6,000 and the rabbit IgG reacted strongly with OMP with the molecular weights of 40,000, 35,000, and 6,000 from the bacteria including Salmonella which belongs to Enterobacteriaceae. The IgY and rabbit IgG also reacted with other proteins from Salmonella typhimurium in immunoblot assay. Competitive ELISA showed that IgY showed specifity to react with two strains of Salmonella typhimurium and Salmonella cholerasuis but not with Escherichia coli and Yersinia enterocolitica. Two strains of Salmonella typhimurium added to UHT milk showed the highest absorbance of all the bacteria used in the sandwich ELISA. Some strains of Salmonella cholerasuis showed higher absorbances than non-Salmonella bacteria.