• Title, Summary, Keyword: ELISA

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Applicability of ABC-ELISA and Protein A-ELISA in serological diagnosis of cysticercosis (유구낭미충증의 혈청학적 진단을 위한 ABC-ELISA와 Protein A-ELISA의 유용성)

  • Lee, Jong-Hyun;Kong, Yoon;Ryu, Jae-Young;Cho, Seung-Yull
    • The Korean Journal of Parasitology
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    • v.31 no.1
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    • pp.49-56
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    • 1993
  • Specific antibody test in serum and cerebrosinal fluid (CSF) is still the main mode of serological diagnosis of cystiercosis. Of different techniques of artibody test, enzyme-linked immunosorbent assay (micro-ELISA) has widely been applied. This study was undertaken to observe whether diagnostic capability can be Improved by applying more sensitive techniques such as Protein A-ELISA and avidin biotin complex ELISA (ABC-ELISA). When evaluated using 115 sera of human cysticercosis, the antibody positive rates were not significantly improved in Protein A-ELISA (82.6%) and in ABC-ELISA (86.1%) than in micro-ELISA (81.7%). The specificities, evaluated in 165 sera from other diseases and normal controls, were significantly improved (88.5% by micro-ELISA, 93.3% by Protein A-ELISA and 93.8% by ABC-ELISA). Antibody levels (absorbance, abs.) in individual serum were correlated well (r : 0.83∼0.86) each other. An actual benefit of Protein A-ELISA and ABC-ELISA was that they needed smaller amount of test sample.

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Studies on Enzyme-linked Immunosorbent Assay(ELISA) for Detection of Antibody to Mycobacterium bovis in Serum and Milk (Enzyme-linked Immunosorbent Assay(ELISA)를 이용한 혈청 및 원유 중의 Mycobacterium bovis 항체 검출에 관한 연구)

  • 심항섭;국정희;박병옥;김성열;박유순
    • Korean Journal of Veterinary Service
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    • v.20 no.2
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    • pp.133-142
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    • 1997
  • In order to supplement a diagnostic method for detection of infectious cattle to bovine tuberculosis, performed ELISA for detection of antibody to if bovis in serum and milk. The diagnostic efficacy of the established ELISA was compared with test of the tuberculin skin test for bovine tuberculosis. The positive corresponding rate of serum ELISA and tuberculin skin test showed 84.3%, milk ELISA and tuberculin skin test showed 75.0%, milk ELISA and serum ELISA showed 75.0% respectively. Comparison of the serum and milk to tuberculin antibody concentration in tuberculin positive cattle, the milk contained 1/100-1/150 concentration compared serum tuberculin concentration. The established ELISA was considered efficient for detection of antibodies to M bovis in serum and milk.

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Comparative evaluation of two commercial ELISA kits for detection of PRRS antibodies using sera collected from pigs in various stages of PRRSV infection (다양한 PRRSV 감염상태에 있는 돼지 혈청을 이용한 PRRS 항체 ELISA 키트들의 비교 평가)

  • Seo, Byoung-Joo;Kim, Hyoun-Il;Kim, Won-Il
    • Korean Journal of Veterinary Service
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    • v.37 no.3
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    • pp.151-156
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    • 2014
  • Porcine reproductive and respiratory syndrome virus (PRRSV) causes major economic losses to the Korean pig industry. ELISA tests using recombinant nucleocapsid protein of PRRSV have been most commonly used for PRRS diagnostics. In the current study, two commercial PRRSV ELISA kits (Bionote PRRSV Antibody ELISA and IDEXX 3XR PRRS Antibody ELISA) have been compared using sera collected from 19 swine farms in various stages of PRRSV infection confirmed by professional diagnostic centers. Thus 130 sera collected from 5 different farms with active PRRSV infection, 130 sera from 6 different farms with PRRS-stabilized status, and 140 sera from 8 different farms with PRRS-free status were evaluated to determine the correlation of test results between those ELISA kits. Both ELISA kits showed a good correlation [PRRSV-positive farms ($R^2$=0.6375) and stabilized farms ($R^2$=0.8928)] in sample-to-positive (S/P) ratio va lues. Among the 140 sera from negative farms, one sample was falsely positive by either of the ELISA kits. In conclusion, both of the ELISA kits showed a good correlation when applied on field samples collected from farms at various stages of PRRSV infection. Bionote ELISA or IDEXX ELISA gave a false positive result on 1 out of 140 negative samples so their specificity was calculated as 99.3%. Therefore, Bionote ELISA would be a good complementary and alternative method for IDEXX ELISA kit, and vice versa.

$F(ab)_2$-ELISA for the Detection of Nuclear Polyhedrosis Virus of Silk-worm, Bombyx mori L.

  • Sivaprasad, V.;Nataraju, B.;Baig, M.;Samson, M.V.;Datta, R.K.
    • International Journal of Industrial Entomology
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    • v.6 no.2
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    • pp.179-181
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    • 2003
  • $F(ab`)_2$-ELISA and direct antigen coating-ELISA (DAC-ELISA) were evaluated in the detection of purified Bombyx mori nuclear polyhedrosis virus (BmNPV) and nuclear polyhedrosis virus infection in silkworm larvae inoculated with BmNPV polyhedra. Although nanogram levels of BmNPV was detected in both DAC- and $F(ab`)_2$-ELISA, similar concentrations of antigen was detected in case of F(ab’)$_2$-ELISA even at higher dilution of antibody (up to 1 : 20 K). One hundred percent nuclear polyhedrosis infection was detected 6 hrs after inoculation in BmNPV infected silkworm larvae by $F(ab`)_2$-ELISA. On the other hand, detection of 100% infection was observed only three days after inoculation in DAC-ELISA. In this study, it was observed $F(ab`)_2$-ELISA was more sensitive than DAC-ELISA in the detection of purified BmNPV as well as nuclear polyhedrosis infection in silkworm larvae.

Nonspecific Mouse Hepatitis Virus Positivity of Genetically Engineered Mice Determined by ELISA

  • Han, Dae Jong;Kim, Hyuncheol;Yeom, Su-Cheong
    • Biomedical Science Letters
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    • v.21 no.1
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    • pp.9-14
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    • 2015
  • Mouse hepatitis virus (MHV) is a major pathogen in laboratory mice that usually leads to fatal diseases, such as hepatitis, multiple sclerosis, encephalitis, and respiratory disease. MHV has a high infection rate, and it needs to be detected as soon as possible to prevent its spread to other facilities. However, MHV detection by enzyme-linked immunosorbent assay (ELISA) often gives false positives; thus, it is very important that the results are confirmed as true positives in the early infection stage or distinguished as false positives with more accurate, reliable methods. Under microbiological screening, MHV ELISA-positive mice were found in four GFP-tagging transgenic mice. To verify the detection of the MHV antigen directly, reverse transcription polymerase chain reaction (RT-PCR) was performed, and the mice were determined to be MHV negative. Additional serum antibody-based screening was conducted with three different ELISA kits, and multiplexed fluorometric immunoassay (MFIA) was performed to confirm their accuracy/sensitivity. In brief, the ELISA kit for A59 nucleocapsid protein (MHV-A59N) revealed MHV ELISA positivity, while other ELISA kits (MHV-S lysate and MHV-JHM lysate) demonstrated MHV negativity. In MFIA, only the test for the recombinant A59 nucleocapsid antigen was MHV positive, which was consistent with the ELISA results. These results suggest that the ELISA kit with the recombinant A59 nucleocapsid antigen might induce non-specific MHV ELISA positivity and that confirmation is therefore essential.

A comparative study on the diagnosis of ELISA test and PPD test of the bovine tuberculosis (ELISA를 이용한 우결핵검사 결과에 대한 PPD 접종법 결과 분석 비교)

  • Lee, Jong-Jin;Kim, Duk-Soon;Lee, Jong-Hwa;Lee, Cheong-San
    • Korean Journal of Veterinary Service
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    • v.33 no.4
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    • pp.335-340
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    • 2010
  • On the basis of the 2009 business plan, 20,394 Korean native cattle and beef cattle were carried examination of bovine tuberculosis by using ELISA technique from March to December. As a result, 66 cattle tested positive for tuberculosis and showed 0.32% positive ratio. Intradermal tuberculin test about 66 cases of ELISA positive cattle was carried out, and all of 66 cattle were confirmed as negative. However, when 7 PPD-positive cattle derived from slaughterhouse were tested by 20k ELISA kit and MS ELISA kit, 3 (2 suspect) cattle and 5 cattle showed positive results, respectively. As compared to the results of PPD test, the concordance rates were 43% (71% included suspect) with 20k ELISA kit and 71% with MS ELISA kit.

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
    • The Korean Journal of Parasitology
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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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Application of Competitive ELISA Method for Estimation of Urinary Aflatoxin M1 Level (ELISA 방법을 이용한 요중 아플라톡신 M1 측정)

  • Kim, Yong-Dae;Kim, Heon
    • Journal of Life Science
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    • v.23 no.2
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    • pp.306-310
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    • 2013
  • We compared the efficacy of the competitive ELISA method for measuring the level of urinary aflatoxin M1 (AFM1) with that of the HPLC-fluorescence detector (HPLC-FLD) method. The recovery rate of AFM1 with the ELISA method was 105% (73-124%), and the coefficient of variation of the analysis was 6.85%. The ELISA method showed a 0.20 pg/ml and 0.62 pg/ml limit of detection and limit of quantitation, respectively. In correlation analysis, the two methods showed a very strong and statistically significant correlation (R=0.96, p<0.01). However, in spite of the strong correlation, the ELISA method tended to overestimate the urinary AFM1 concentration compared to the HPLC-FLD method. These results suggest that the competitive ELISA method may be a useful technique for measuring the AFM1 level in high-throughput urine samples, but it needs to be corrected with a regression equation from regression analysis with the HPLC-FLD method.

Comparison of serological methods for detection of avian influenza virus antibodies (가금인플루엔자 바이러스 항체검출을 위한 혈청학적 진단법 비교)

  • Han, Myung-guk;Park, Kyoung-yoon;Kwon, Yong-kuk;Kim, Jae-hong
    • Korean Journal of Veterinary Research
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    • v.42 no.1
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    • pp.73-80
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    • 2002
  • An enzyme-linked immunosorbent assay (ELISA) using purified hemagglutinin of swine influenza virus (H1N1) as antigen was developed for detection of antibody to avian influenza virus (AIV). The sensitivity and specificity of a developed and commercial available ELISA kits were compared with those of agar gel precipitation (AGP) test and hemagglutination inhibition (HI) test using sera collected from chickens under condition of field exposure. The concentration of antigen, serum dilution and concentration of enzyme-conjugated secondary antibody in developed ELISA (S-ELISA) were 0.5ug/100ul, 1:200 and 0.03ug/100ul, respectively. The correlation coefficients between S-ELISA and commercial ELISA and HI titers were 0.419 and 0.533, respectively. A significant correlation (p < 0.01) was not found between HI and ELISA titers. The S-ELISA was found to be as more sensitive and specific than the AGP test, showing 86.8% sensitivity and 85.3% specificity. It is suggested that the ELISA using the SIV as antigen may be useful method as an investigating tool for AIV serological surveillance.

Evaluation of the extract from lymph nodes for Salmonella monitoring in pig herds (돈군의 Salmonella 모니터링을 위한 림프절 추출액 사용에 대한 평가)

  • Jung, Byeong-Yeal;Choo, Ji-Hoon;Kim, Ji-Hun;Jung, Jae-Yun
    • Korean Journal of Veterinary Research
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    • v.46 no.2
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    • pp.119-125
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    • 2006
  • The objective of this study was to investigate the use of extract from mesenteric lymph nodes as an alternative to serum for ELISA to detect Salmonella antibodies in slaughter pigs. Among 324 slaughter pigs, 65 (20.1 %) were positive in the serum ELISA and 76 (23.5%) were positive in the ELISA with extract from lymph nodes. A total of 24 (7.4%) Salmonella representing 6 serotypes were isolated from mesenteric lymph nodes and 35 (10.8%) Salmonella belonging to 2 serotypes were also recovered from cecal contents of slaughter pig samples, respectively. The most prevalent serogroup was B (55.9% of isolates) and serotype was Typhimurium (52.5% of isolates). In the comparison of the results of between the serum ELISA and Salmonella isolation, kappa value was 0.28 with mesenteric lymph nodes and 0.37 with cecal contents, respectively. However, the extract ELISA had sensitivity of 98.5%, specificity of 95.4% and kappa value of 0.88 as compared with the serum ELISA. Because high degree of concordance between the serum ELISA and the extract ELISA was observed (P=0.24), extract from lymph nodes could be used as an alternative to serum for the detection of Salmonella antibodies in the ELISA.