• Title/Summary/Keyword: antibody detection

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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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Serum Antibody Ig G and Ig M Titers for Opisthorchis felineus Correlate with Eggs in Faeces - a Comprehensive Study in Chuvash Republic, Russia

  • Emelianov, Vladimir U;Skvortsova, Tatiana;Mikhailova, Lilia V;Shamitova, Elena N
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.1
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    • pp.281-283
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    • 2016
  • The Cholangiocarcinoma is a. The risk of development of cholangiocarcinoma, generally a rare type of a liver tumor, increases during infection of Opisthorchiasis. For this reason the timely detection of Opisthorchiasis is important for Cholangiocarcinoma prevention. There are many studies which concern the detection of pathogenesis of Opisthorchis viverrini infection but a little known about Opisthorchis felineus. In this study we investigate a correlation of the eggs which are found in a faeces and are comparable with a serum Ig G and Ig M antibody level that were detected with ELISA test in a large group of patients. The result is showing positive correlation between evidence of the Opisthorchis felineus eggs that were found in a faeces and antibody Ig G and Ig M level in a serum. Moreover the combination of two methods can improve the Opisthorchiasis diagnostic: the serum antibody and faeces investigation of eggs.

Immunoassay of haptoglobin and transferrin with proteinG-containing QCM sensor chip and unpurified antiserum (Protein G를 포함하는 수정미소저울 센서 칩과 정제되지 않은 항혈청을 이용한 헵토글로빈과 트랜스페린의 면역분석)

  • Ha, In-Young;Choi, Suk-Jung
    • Journal of Sensor Science and Technology
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    • v.17 no.5
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    • pp.380-386
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    • 2008
  • Quartz crystal microbalance immunosensor has a capacity to perform a label-free and real time detection of a trace amount of analyte through the specific interaction between antibody and antigen. However, immobilization of antibody molecules on the sensor surface is a troublesome procedure for researchers who are not experienced in chemistry. Protein G has a specific affinity to antibody and would serve as a capturing agent for antibody when immobilized on the sensor surface. In this work, we prepared a protein G sensor chip by immobilizing protein G on the surface of quartz crystal microbalance and examined its capability to detect human haptoglobin or human transferrin with unpurified corresponding antiserum. Specific and dose dependent response was observed when the protein G chip was used for detection of antigens after saturated with antiserum. We also verified several advantageous aspects of the protein G chip such as improved flexibility and sensitivity.

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.

Prevelance of neutralizing antibody related with viral respiratory disease in cattle (송아지 바이러스성 호흡기 질병 중화항체 조사)

  • Youn, Choong-Keun;Lim, Yeon-Soo;Lyoo, Young S.
    • Korean Journal of Veterinary Research
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    • v.50 no.3
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    • pp.205-211
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    • 2010
  • Fifty young calves, about five to six months old purchased from nation-wide were investigated with the prevelance of neutralizing antibody (Ab) of infectious bovine rhinotracheitis virus (IBRV), parainfluenza 3 virus ($PI_{3}V$), bovine viral diarrhea virus (BVDV) and bovine respiratory syncytial virus (BRSV). The positive detection ratio of neutralizing Ab against IBRV was only 3% and two of positive samples showed low antibody titer (below 2). Ab against BRSV showed 48% of positive ratio and among 24 positive samples, antibody titer of 23 samples were below 3. But in the case of BVDV, 68% of samples were positive and 23 samples appeared to possess high antibody titer, above 4 and the antibody titer of five samples were above 8. The highest positive result came from $PI_{3}V$. The positive ratio in the samples investigated in this study was 72%, but the antibody titer of positive samples were generally below 3 (77.8% in positive samples).

Studies on the RBC Alloimmunization after Blood Transfusions

  • Kim Jae-Woo;Kim We-Jong
    • Biomedical Science Letters
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    • v.12 no.1
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    • pp.35-42
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    • 2006
  • Alloimmunization to red blood cell (RBC) antigens may cause a delayed hemolytic transfusion reactions (DHTR) and a delayed serologic transfusion reactions (DSTR). In the present study, the frequency of alloimmunization and its clinical significance were evaluated. Also, transfusions were correlated with antibody formation. Alloimmunization rate was 0.63%. Alloimmunization rate in multiple transfused patients was 24.5%. The most common clinically significant alloantibodies of alloimmunized patients were found to be Rh antibodies (52.6%). Nine patients out of 38 (23.7%) became undetectable after the first detection. To be positive at antibody screening test after RBC transfusion was mean transfused numbers: 3.7 units, mean transfused periods: 56 days, mean transfused frequencies: 1.7 times. The results from antibody specificity and RBC transfusions were comparatively analyzed and it shows that Rh system antibodies were longer than other antibodies (P<0.05). In case of disease group, malignant diseases was longer than other diseases (P<0.05). In order to prevent the formation of RBC alloimmunization, irregular antibody screening tests were performed at propriety intervals in multiple transfused patients.

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Evaluation of Antibody Immobilization Methods for Detection of Salmonella using Impedimetric Biosensor (살모넬라균 검출을 위한 임피던스 바이오센서의 항체 고정화 방법 평가)

  • Kim, Gi-Young;Moon, Ji-Hea;Om, Ae-Son;Yang, Gil-Mo;Moh, Chang-Yeon;Kang, Suk-Won;Cho, Han-Keun
    • Journal of Biosystems Engineering
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    • v.34 no.4
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    • pp.254-259
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    • 2009
  • Conventional methods for pathogen detection and identification are labor-intensive and take several days to complete. Recently developed biosensors have shown potential for the rapid detection of foodborne pathogens. In this study, an impedimetric biosensor was developed for rapid detection of Salmonella typhimurium. To develop the biosensor, an interdigitated microelectrode (IME) was fabricated by using semiconductor fabrication process. Anti-Salmonella antibodies were immobilized based on either avidin-biotin binding or self assembled monolayer (SAM) on the surface of the IME to form an active sensing layer. To evaluate effect of antibody immobilization methods on sensitivity of the sensor, detection limit of the biosensor was analyzed with Salmonella samples innoculated in phosphate buffered saline (PBS) or food extract. The impedimetric biosensor based on SAM immobilization method produced better detection limit. The biosensor could detect 107 CFU/mL of Salmonella in pork meat extract. This method may provide a simple, rapid, and sensitive method to detect foodborne pathogens.

Development of cellulose nano beads based a rapid detection kit to detect staphylococcal enterotoxin B

  • Kim, Giyoung;Yoo, Jinyoung;Park, Saetbyeol
    • Korean Journal of Agricultural Science
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    • v.46 no.3
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    • pp.549-557
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    • 2019
  • Staphylococcal enterotoxin is a very common cause of food poisoning. Conventional detection methods for the toxin including enzyme-linked immunosorbent assays (ELISAs), chemiluminescence (ECL), and polymerase chain reaction (PCR) assays require a lot of time, efforts, and expert technicians. Lateral flow strip kits have shown great potential for the rapid detection of foodborne pathogens. The lateral flow strip kit is widely used in clinical settings because it is easy to use, fast, and cost effective. A typical lateral flow strip kit uses colloidal gold to generate a visual signal. However, the lateral flow strip kit based on colloidal gold has limited sensitivity to fulfill food safety regulation requirements. This study was performed to develop a rapid test kit for pathogenic staphylococcal enterotoxin B (SEB) in food samples. The rapid detection kit was fabricated based on a nitrocellulose lateral-flow strip. Cellulose nano beads and SEB antibodies were used as the tag and receptor, respectively, to improve the detection performance. Manually spotted SEB antibody and anti-rabbit antibody on the surface of the nitrocellulose membrane were used as test and control spots, respectively. The feasibility of the rapid test kit to detect SEB in samples was evaluated. The sensitivity of the kit was 10 ng/mL SEB spiked in PBS. Additionally, the rapid test kit could detect 1 ng/mL of SEB in chicken meat extract.

Antibody Layer Fabrication for Protein Chip to Detect E. coli O157:H7, Using Microcontact Printing Technique

  • KIM HUN-SOO;BAE YOUNG-MIN;KIM YOUNG-KEE;OH BYUNG-KEUN;CHOI JEONG-WOO
    • Journal of Microbiology and Biotechnology
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    • v.16 no.1
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    • pp.141-144
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    • 2006
  • An antibody layer was fabricated to detect Escherichia coli O157:H7. The micropattern of 16-mercaptohexadecanoic acid (16-MHDA) as alkylthiolate was formed on the gold surface by using the PDMS stamp with microcontact printing $({\mu}CP)$ techniques. In order to form antibody patterns on the template, protein G was chemically bound to the 16-MHDA patterns, and antibody was adsorbed on a self-assembled protein G layer. The formation of the 16-MHDA micropattern, self-assembled protein G layer and antibody pattern on Au substrate was confirmed by surface plasmon resonance (SPR) spectroscopy. Finally, the micropatterning method was applied to fabricate the antibody probe for detection of E. coli O157:H7, and monitoring of antigen by using this probe was successfully achieved.

Recognition of Microorganisms Using SPR Biosensor Immobilized with Thiolated Antibody (티올화 항체고정형 SPR 바이오센서를 이용한 미생물 인식)

  • 조용진;김남수
    • Journal of Biosystems Engineering
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    • v.28 no.2
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    • pp.167-172
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    • 2003
  • This study was performed to fabricate a batch-type SPR biosensing system using a thiolated E. coli antibody coupling, and to explore the feasibility of real-time detection of E. coii in a stagnant sample solution. In advance. “O” and “K” antigenic serotype E. coli antibodies were thiolated with sulfo-LC-SPDP and dithiothreitol, and immobilized by chemisorption in the gold surface of compact SPR sensors. When the SPR biosensor immobilized with E. coli antibody monitored a E. coli solution, it took 3 to 5 min to stabilize. The SPR biosensing system developed in this study was able to detect E. coli in the range above 10$^4$ CFU/mL at the 0.05 significant level. Also, the SPR biosensor had possibility to significantly detect E. coli in the range of 10$^2$ to 10$^4$ CFU/mL in E. coli solutions. Meanwhile, when the SPR biosensor immobilized with 5. coli antibody was cleaned with NaOH solutions, its ability to detect E. coli largely decreased due to wash-out of the immobilized antibody. In order to reuse the SPR sensor, it should be antibody-immobilized newly.