• Title/Summary/Keyword: ELISA antibody

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Studies on enzyme-linked immunosorbent assay(ELISA) for detection of antibody to Brucella abortus (효소면역법을 이용한 Brucella abortus 항체 검출에 관한 연구)

  • 심항섭;국정희;정봉수;고태오;조중현;박유순
    • Korean Journal of Veterinary Service
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    • v.21 no.2
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    • pp.107-115
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    • 1998
  • In order to establish a rapid, sensitive and specific diagnostic method for detection of antibody to Brucella abortus, a enzyme-linked immunosorbent assay(ELISA) was adapted. The diagnostic efficacy of the established ELISA was compared with that of the standard tube agglutination test for B abortus. 1. It was found that the optimal concentration of antigen for this ELISA was 5$\mu\textrm{g}$/ml, the optimal dilution of conjugate was 1 : 2000, and the optimal dilution of serum was 1 : 200, respectively. 2. Cut off value in this ELISA was 1,102 that was determined by mean absorbance(at 492nm) of tube agglutination test negative serum added with the triple value of the standared devation. 3. The relationship between the tube agglutination test and ELISA was showen high corresponding rate with sensitivity(96.3%) and specificity(98.1%). 4. The efficacy of the ELISA for detection of B abortus antibody was compared with tube agglutination test In brucellosis outbreak farm. The sensivity of ELSIA was higher than tube agglutination test.

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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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Development of monoclonal antibody capture ELISA for the detection of antibodies against transmissible gastroenteritis virus

  • Oh, Yeonsu;Tark, Dongseob
    • Korean Journal of Veterinary Service
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    • v.42 no.1
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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.

Studies on the combined inactivated oil emulsion vaccine of Newcastle disease and avian infectious bronchitis in chickens (닭의 뉴캣슬·전염성 기관지염 바이러스 혼합 불활화 오일 에멀션 백신의 생산시험)

  • Jeon, Yun-seong;Kim, Sun-joong;Seo, Ik-soo
    • Korean Journal of Veterinary Research
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    • v.30 no.1
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    • pp.79-84
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    • 1990
  • A single inoculation of combined vaccines of Newcastle disease and avian infectious bronchitis of chicken, in a form of gel-oil emulsion type (gel-OEV) was tested their immunogenecity in chickens. The results were summerized as follows: 1. Average minimum and maximum ELISA antibody titers of ND were recorded 2407 and 13144 respectively. In the case of IB, 1824 and 4496 were recorded as minimum and maximum titers. 2. The distribution of average proportional groups, in the lowest and the highest, were 1.6 and 7.0 in ND ELISA and 1.4 and 2.8 in IB ELISA antibody titers. 3. ND ELISA antibody titers were significantly increased upto 7th week after the vaccination. On the other hand, IB ELISA antibody titers were raised upto 4th week after the vaccination.

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Study on Development of Enzyme-Linked Immunosorbent Assay for the Screening of Chloramphenicol Residues (잔류 Chloramphenicol 검사용 효소 면역측정법의 개발에 관한 연구)

  • 윤동호;이문한
    • Journal of Food Hygiene and Safety
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    • v.8 no.4
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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.

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Changes of maternal antibodies in chicks vaccinated to breeder against infectious bronchitis, infectious Bursal disease, and Newcastle disease virus (모계의 전염성기관지염, 전염성 F낭병 및 뉴캣슬병 백신투여에 따른 모체이행 항체의 변동)

  • 고원석;김태중;이정원;서이원;송희종;오언평
    • Korean Journal of Veterinary Service
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    • v.21 no.2
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    • pp.133-139
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    • 1998
  • Serum samples collected from 30 breeders and their progeny 30 chicks. The antibodies against infectious bronchitis(IB), infectious bursal disease (IBD) and Newcastle disease(ND) viruses were detected by ELISA using commercial ELISA kit. The breeders were vaccinated against IB, IBD and ND viruses according to general vaccination program. Geometric mean titers(GMT) of ELISA were monitored from 1-day old to 17-day old chicks and compared with breeder chickens. The GMT of ELISA to IB, IBD and ND were declined half level of the breeder antibody titer at 6-, 8- and 7-day old. And, the GMT of ELISA to IB, IBD and ND were declined than that of protective titer at 6-, 1-, and 4-day old. Thereafter, the GMT of ELISA was declined and disappeared according to ages of chicks. Taken together, this study led to conclusion that time-course of maternal antibody titers of chicks from vaccinated breeders, and this is very important data for vaccination to chicks.

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Production of the Monoclonal Antibodies to the Escherichia coli Heat-Stable Enterotoxin (대장균의 내열성장독소 측정법개발을 위한 단세포군항체의 생산)

  • Chang, Woo-Hyun;Lee, Woo-Kon;Kim, Suck-Yong;Park, Jung-Bum
    • The Journal of the Korean Society for Microbiology
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    • v.22 no.4
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    • pp.377-392
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    • 1987
  • Monoclonal antibody to the Escherichia coli heat-stable enterotoxin(ST) was produced to develop a rapid and convenient diagnostic method to the ST. The toxin was purified from culture supernatant of enterotoxigenic E. coli O148H28($ST^+/LT^+$) and conjugated to bovine serum albumin(BSA). The ST-BSA conjugate was used to immunize BALB/c mice and the immune spleen cells from these mice were fused with $P3{\times}63$ Ag8.V653 plasmacytoma cells. Hybridomas were screened by ELISA and positive hybridomas were cloned by limiting dilution. Finally, one stable clone (AS36) specific to ST was selected for further growth and characterization. Antibody titers of culture supernatant and ascitic fluid from BALB/c mice were 1:1,024 and 1:20,480 respectively in ELISA. The isotype and subclass of monoclonal antibody was IgG1 in sandwich ELISA. To test the neutralizing effect of monoclonal antibody on toxin activity of ST, mixture of ascitic fluid and ST was assayed by infant mouse assay and this monoclonel antibody was proved to be a neutralizing antibody. The titer of ascitic fluid which completely neutralized biological activity of 4 units of ST was 1:4. Purified ST was quantitatively measured by competitive ELISA and minimum amount of ST detectable by this assay was 250pg, which was an amount six-fold smaller than that detectable by infant mouse assay. Four reference strains of enterotoxigenic E. coli from WHO were detected by competitive ELISA and highly specific, sensitive and reproducible result was obtained.

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Detection of Antibodies in Serum and Cerebrospinal Fluid to Tonoplasma gondii by Indirect Latex Agglutination Test and Enzyme-Linked Immunosorbent Assay (간접 Latex 응집반응과 ELISA에 의한 중추신경계 질환 환자의 혈청 및 뇌척수액에서 Toxoplasmu gondii에 대한 항체 검출)

  • 최원영;남호우
    • Parasites, Hosts and Diseases
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    • v.30 no.2
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    • pp.83-90
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    • 1992
  • Sensitivity of anti-Texoplasma antibody (IgG) test by enzyme·linked immunosorbent assay (ELISA) was evaluated in comparison with indirect laten agglutination (ILA) using 2,016 paired human samples of serum and cerebrospinal quid (CSF) . The samples were collected from neurologic patients in Korea with mass lesions in central nervous system(CNS) as revealed by imaging diagnosis(CTIMRI). When the sera were screened for anti-Toxoplasma antibody by ILA, 76 cases (3.8%) were positive (1:32 or higher titers). In the pairs samples of CSF, no positive reactions were observed. When ELISA was performed using PBS extract of Percoll purified tachysoites as antigen, cut-off absorbance was determined as 0.40 for serum and 0.27 for CSF tests. The antibody positive rates by ELISA were 7.0% in serum and 5.6% in CSF Of them, 40 cases(2.0%) showed positive reactions in both serum and CSF, The antibody positive rates were higher in groups older than 40 years, The rates were higher in male(4.7% by ILA, 8.3% by ELISA) than in female(2.2% by ILA, 5.0% by ELISA). The rates in CSF showed no such sex difference. ELISA showed twice higher positive rates when serum was tested, and was sensitive enough to detect specific antibodies in CSF. Etiologic relations between positive antibody tests and CNS lesions remained unknown.

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Studies on serological tests for pullorum disease (추백리의 혈청학적 진단법에 관한 연구)

  • 김정태;심항섭;김태종;고태오;우종태;유기승;박유순
    • Korean Journal of Veterinary Service
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    • v.21 no.3
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    • pp.313-323
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    • 1998
  • In order to establish a sensitive and specific diagnostic method for detection of antibody to Salmonella pullorum, a enzyme-linked immunosorbent assay(ELISA) was designed and standardized. The diagnostic efficacy of the established ELISA was compared with that of the serum plate agglutination test and immunodiffusion test for pullorum disease. 1. The chicken hyperimmune sera to Salmonella pullorum, S gallinarum, S typhimurium and S typhi were shown the cross reaction to S pullorum antigen by serum plate agglutination test. 2. When compared the cross reaction titer of microplate agglutination test for chickens hyperimmune sera, it was found that the titer were 64 in S pullorum, 32 in S gallinarum, 4 in S typhimurium and 8 in S typhi, respectively. 3. When compared the specificity of various antigen(HA, EA, PA and SA) by the immunodiffusion test, the most suitable antigen was phenol-treated bactrium. 4. The optimal concentration of S pullorum antigen for ELISA was 1 : 160 dilution of bacterium. 5. The efficacy of the ELISA for detection of S pullorum antibody was compared with serum Plate agglutination test and immunodiffusion test in chickens infected with S pullorum. The antibody was first detected at 6 days after infection using three tests examined. The antibody was alldetected at 9 days by ELISA, at 12 days by serumplate agglutination test, at 15 days by immunodiffusion test.

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Quantitation of Recombinant Hirudin by Enzyme-Linked Immunosorbent Assay (효소면역측정법 (ELISA)을 이용한 유전자 재조합 히루딘의 정량)

  • Choi, Yun-Joo;Hahn, Bum-Soo;Ahn, Mi-Young;Park, Pyung-Keun;Sohn, Jung-Hoon;Choi, Eui-Sung;Lee, Sang-Ki;Kim, Yeong-Shik
    • YAKHAK HOEJI
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    • v.41 no.1
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    • pp.74-80
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    • 1997
  • A polygonal antibody against recombinant hirudin was raised for the development of a ELISA in biological fluids. Recombinant hirudin was conjugated to maleimide activated carrie r protein, KLH and injected to a rabbit. The third booster collection of antiserum was used as primary antibody for the ELISA. The titer for the detection antibody was determined. The direct ELISA could determine the concentration of hirudin in the range of ~10ng/ml. Affinity pulified IgG was obtained and conjugated to horseradish peroxidase. Purified IgG and IgG-HRP could be used as capture and detection antibody, respectively. Although sandwich ELISA would not give the satisfactory results. it could apply for the detection of hirudin level in the range of ~20 ${\mu}$g/ml.

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