• Title/Summary/Keyword: ELISA

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효소면역흡착시험을 이용한 경북서부지역의 돼지 흉막폐렴에 대한 항체분포조사 (Survey on the distributions of swine pleuropneumonia antibodies by ELISA in Kyongbuk western area)

  • 서희진;배성수;김대원;김봉환
    • 한국가축위생학회지
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    • 제23권3호
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    • pp.289-299
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    • 2000
  • The study was performed to investigate the distributions of swine pleuropneumonia in Kyongbuk western area by the enzyme-linked immunosorbent assay (ELISA). Sera collected from 400 slaughtered pigs in 3 slaughter houses during the period from May 1999 to october 1999 were tested to detect antibodies against A pleuropneumonie serotype 2 and 5. The optimal dilution of CBE antigen, conjugate and serum for this ELISA were determined 1 : 400, 1 : 20,000, 1 100, respectively. The optimal dilution of OW antigen, conjugate and serum for this ELISA were determined 1 : In, 1 : 20,000, 1 : 200, respectively. Cut-off value in this ELISA was determined by mean absorbance (at 492 nm) of negative control sera added with the triple value of the standard deviation. Cut-off value in ELISA by CBE and OMP antigen were 1.134 and 1.217, respectively. By the ELISA, positive reaction rates to A pleuropneumoniae serotype 2 and 5 for CBE antigen were 38.8% and 18.8%, and for OMP antigen were 42.8% and 23.5% of the 400 samples, respectively.

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

  • 이승배;최석호;최재원
    • 한국축산식품학회지
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    • 제24권2호
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    • pp.182-188
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    • 2004
  • Lactoperoxidase(LPO)를 농도를 측정하기 위한 ELISA을 개발하기 위해 LPO로 면역시킨 갈색 산란계의 계란에서 형성된 anti-LPO IgY 항체를 분리 정제하고, 분리된 anti-LPO IgY 항체의 특이성을 ELISA 와 double immunodiffusion 방법으로 조사한 후 indirect ELISA 방법을 이용한 표준곡선을 만들었다. 분리 정제된 anti-LPO IgY항체의 titer는 1:520,000이며, ELISA와 double immunodiffusion 방법 모두에서 $\alpha$-lactalbumin, $\beta$-lactoglobulin, casein 및 lysozyme하고는 교차반응을 하지 않고 LPO만 높은 특이성을 갖는 항체로 나타났다. Indirect ELISA방법에서 LPO의 coating 농도는 0.25 $\mu\textrm{g}$/mL이며 anti-LPO IgY 최적 희석배수는 1:8,000으로 나타났다. Indirect ELISA 방법으로 LPO를 측정할 수 있는 표준곡선에서 민감도의 범위는 0.0l-l $\mu\textrm{g}$/mL로 나타났다.

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

  • 고원석;이정원;곽길한;권정택;송희종
    • 한국가축위생학회지
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    • 제24권1호
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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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육계의 뉴켓슬병 방어역가 측정에 있어서 ELISA 검사법의 효용성 (Efficacy of ELISA for measurement of protective newcastle disease antibody level in broilers)

  • 김종녀;허원;모인필
    • 대한수의학회지
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    • 제46권3호
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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.

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
    • Parasites, Hosts and Diseases
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    • 제41권1호
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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.

육계에서 전염성기관지염, 전염성 F 낭병, 뉴캣슬병 백신투여에 따른 혈중항체가의 변동 (Changes of maternal antibodies in broilers vaccinated with infectious bronchitis, infectious bursal disease and Newcastle disease viruses detected by ELISA)

  • 고원석;백귀정;이정원;서이원;김태중;송희종;오언평
    • 한국가축위생학회지
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    • 제21권3호
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    • pp.277-284
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    • 1998
  • Serum samples were collected from 100 breeders and their progeny 600 broilers. The breeders and broilers were vaccinated against infectious bronchitis(IB), infectious bursal disease(IBD) and Newcastle disease(ND) viruses according to general vaccination program. The antibodies in serum samples against IB, IBD and ND viruses were detected by ELISA using commercial ELISA kit. Geometric mean titer(GMT) of ELISA was monitored from 1-day-old to 35-day-old broilers and compared to that of breeder chickens. The GMT of ELISA to IB, IBD and ND was declined half level of the day old broiler's antibody titers at about 4, 9 and 4 days of age. The GMT of ELISA to IB, IBD and ND was declined than that of protective antibody titer at about 12, 11, and 15 days of age. Thereafter, the GMT of ELISA to IB, ND were declined and disappeared according to age of broilers. The GMT of ELISA to IBD was declined according to age of broilers, but at 25 days of age increased and 31 days of age increased than that of protective antibody titer. Taken together, these studies led to conclusion that time-course of antibody titers of broilers from vaccinated breeders and that of progeny broliers which vaccinated according to vaccine program. Those are very important data to design vaccine program to breeders and broilers.

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Babesia gibsoni의 적혈구내 배양법과 진단법 개발에 관한 연구 1. Babesia gibsoni 진단을 위한 간접형광항체법(IFAT)과 효소표지면역검사법(ELISA) (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)

  • 서명득;신용승
    • 대한수의학회지
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    • 제37권3호
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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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느타리버섯 세균성갈반병균 Pseudomonas tolaasii의 효소면역검출법 (Enzyme-Linked Immunosorbent Assays of Pseudomonas tolaasii, a Bacterial Brown Blotch Pathogen of Oyster Mushroom.)

  • 이향범;전낙범;손동화;유승헌
    • 한국미생물·생명공학회지
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    • 제26권3호
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    • pp.238-243
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    • 1998
  • 느타리버섯의 세균성갈반병균인 P. tolaasii(PT)를 신속 간편하게 검출할 수 있는 효소면역측정법 (ELISA)을 개발하였다. 특이항체를 생산하기 위하여 PT($5{\times}10^7$ cfu)를 Freund's adjuvant와 함께 토끼에 수차례 면역하였으며, 가장 높은 역가를 나타낸 항혈청을 이용하여 비경합 ELISA 및 경합 ELISA를 각각 확립하였다. 표준곡선으로부터 PT의 그 검출한계는 각각 $2{\times}10^2$cfu/ml 및 $3{\times}10^2$cfu/ml가량으로 나타났다. 교차반응은 비경합 ELISA의 경우 P. agarici, P. reactans 및 비병원성 Pseudomonas속 균주와 1/$10^3$이하의 매우 미약한 반응을 보였으나 P. solanacearum, Erwinia chrysanthemi, Streptococcus mutans, Xanthomonas citri 및 곰팡이인 Fusarium oxysporum등 다른 균주와의 반응성이 거의 없었으며, 경합 ELISA의 경우 PT이외에는 같은 속 균주에 대하여도 반응성이 거의 없었다. ELISA를 이용하여 버섯으로부터 분리된 18균주에 대하여 PT의 여부를 조사하였을 때 병원성과 white line 반응성을 동시에 나타내었던 11균주에 대하여만 명확한 양성으로 나타나, 본 연구에서 확립한 ELISA는 간편하고 정확한 PT검출법 임이 확인되었다.

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

  • 최석호
    • 한국축산식품학회지
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    • 제24권2호
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    • pp.176-181
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    • 2004
  • 우유내 Salmonella를 검출하기 위한 Sandwich ELISA의 특이성을 조사하였다. Sandwich ELISA에 사용한 항체들은 OMP 분획을 닭에 면역주사하여 얻은 IgY와 OMP 분획을 gel filtration하여 얻은 분자량 40,000의 OMP를 토끼에 면역 주사하여 얻은 토끼 IgG를 사용하였다. Immnoblot assay에서 IgY는 분자량 6,000의 OMP에 강하게 반응하였으며 토끼 IgG는 분자량 40,000, 35,000과 6,000의 OMP들에 강하게 반응하였다. IgY와 토끼 IgG는 Salmonella typhimurium의 다른 단백질에도 반응하였다. Competitive ELISA에서 IgY가 Salmonella의 두 개 균주에 대해 특이성을 나타냈으며 Eshcherchia coli와 Yersinia enterocolitica에 의하여서는 반응을 나타내지 않았다. 우유에 세균을 첨가하여 실시한 sandwich ELISA에서 Salmonella typhimurium 2균주가 가장 높은 흡광도를 보였다. Salmonella cholerasuis 균주들은 상대적으로 흡광도가 낮았으며 이루 일부 Salmonella cholerasuis 균주들은 비 Salmonella 균주들과 차이가 없었다.

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

  • Han, Dae Jong;Kim, Hyuncheol;Yeom, Su-Cheong
    • 대한의생명과학회지
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    • 제21권1호
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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.