• Title/Summary/Keyword: ELISA IgM

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Enzyme-Linked Immunosorbent Assay(ELISA) for the Rapid Detection of the Flacherie Virus Disease (효소항체법에 의한 누에 바이러스성 무름병의 진단)

  • Gang, Seok-U;Kim, Gwon-Yeong;Gang, Seok-Gwon
    • Journal of Sericultural and Entomological Science
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    • v.34 no.1
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    • pp.35-40
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    • 1992
  • An enzyme-linked immunosorbent assay (ELISA) was studied for the rapid diagnosis of the flacherie virus (FV) of the silkworm, Bombyx mori. The optimised concentration of rabbit anti-FV IgG and enzyme conjugate for the this technique were 15$\mu\textrm{g}$/$m\ell$ and 1:100 dilution, respectively. In ELISA, the detectable concentation of purified FV was 15ng/$m\ell$, and the flacherie viral antigens in the larval extracts were detected as early as 24 hours after the experimental infection. The results indicated that ELISA technique proved to be applicable for the rapid diagnosis of flacherie virus disease.

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A Pneumococcal Conjugate Vaccine Formula Induces Protection in Mice Against Disseminated Disease due to Streptococcus pneumoniae (페렴구균 전신감염에 대한 협막. 표면단백질 접합백신의 효과)

  • Han , Yong-Moon;Lee , Jue-Hee
    • YAKHAK HOEJI
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    • v.48 no.6
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    • pp.345-351
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    • 2004
  • ln the present work to determine effect of a Streptococcus pneumoniae conjugate vaccine, S.pneumoniae capsule attached to the surface protein (JY-Pol) was ex amined. This JY-Pol contained approximately 92% and 6% carbohydrate and protein, respectively. Gel electrophoresis revealed the presence of the surface protein in the JY-Pol. By the double immunodiffusion and isotyping ELISA analyses, administration of JY-Pol that was adsorbed to alum adjuvant (JY-Pol/Alum) into mice induced IgM, IgG, and IgA specific for the S.pneumoniae capsule. The ATCC capsular polysaccharide adsorbed to alum (ATCC-Pol/Alum) provoked only IgM in mice. In survival tests, mice that were immunized with the JY-Pol/Alum before intravenous challenge with live S.pneumoniae survived entire period of 46 day-observation, whereas all mice that received ATCC-Pol/Alum or only diluent instead of the vaccination died within 5 and 12 days, respectively. Results from footpad-edema test showed that JY-Pol/Alum formula provoked the cellular immunity as determined by swelling of the mouse footpad. These data indicate that the naturally conjugated JY- Pol enhances resistance of mice against disseminated pneumococcal disease due to S.pneumoniae by both humoral and cellular immune responses.

Seroprevalence of Toxoplasmosis with ELISA and Rapid Diagnostic Test among Residents in Gyodong-do, Inchon city, Korea: A Four-Year Follow-up

  • Kim, Yeong Hoon;Lee, Ji hoo;Ahn, Seong kyu;Kim, Tong-Soo;Hong, Sung-Jong;Chong, Chom-Kyu;Ahn, Hye-Jin;Nam, Ho-Woo
    • Parasites, Hosts and Diseases
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    • v.55 no.3
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    • pp.247-254
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    • 2017
  • ELISA has been used for the diagnosis of toxoplasmosis, but it is being gradually replaced by a rapid diagnostic test (RDT). We compared and analyzed ELISA and RDT results using the sera collected during 4 consecutive years from residents of Gyodong-do (Island), Incheon-city, Korea. Sera from 921, 993, 940, and 838 adult residents were collected on a yearly basis (2010-2013). ELISA was performed by using a crude extract of T. gondii RH strain antigen and IgG/IgM RDT mounted with recombinant fragment of major surface antigen (SAG1), GST-linker-SAG1A, were applied to the sera. Comparison between groups was analyzed by the Student's t-test. The positive seroprevalence surged from 14.7% (135/921, 2010), 23.1% (231/993, 2011), 23.6% (222/940, 2012), and 32.1% (269/838, 2013) by ELISA. In contrast, RDT showed a more moderate increasing trend from 21.7% (200/921, 2010), 25.5% (253/993, 2011), 28.9% (272/940, 2012) and 33.1% (277/838, 2013). Discrepancies between ELISA and RDT were noted near the cut-off value. At the OD 0.15-0.24 range, RDT could detect 16.1% (169/1051) more positives, which suggests an early or acute toxoplasmosis, but at the OD 0.25-0.34 range, ELISA could detect 35.9% (92/256) more positives of possible chronic infections. Over the OD > 0.35 ELISA and RDT agreed in the majority of the cases. This surge in seroprevalence may be caused by the organic agriculture in addition to eating behavior or increase in pets among Koreans. These facts may be applied on a full-scale national survey using RDT to supplement ELISA to define the characteristics of the infection.

Seroprevalence of Pertussis in Healthcare Workers without Adult Pertussis Vaccine Use at a University Hospital in Korea

  • Choi, Won Suk;Kim, Su Hyun;Park, Dae Won
    • Journal of Korean Medical Science
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    • v.33 no.50
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    • pp.321.1-321.7
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    • 2018
  • Background: Pertussis is highly contagious respiratory disease. Healthcare workers (HCWs) can be an important mediator of the disease. A seroprevalence of pertussis was investigated in HCWs to determine the immune status against pertussis and to detect the unidentified pertussis. Methods: This study was conducted for HCWs at a hospital located in Korea in 2011. After obtaining written informed consent for HCWs voluntarily participating in the study, 10 mL of blood was collected from each subject. Demographic and medical data were collected using questionnaire. Data on the underlying disease and vaccination history were reviewed again through medical records. The presence of anti-pertussis toxin (anti-PT) immunoglobulin G (IgG), and immunoglobulin A (IgA) was detected by quantitative analysis using a commercially available ELISA kit (EUROIMMUN, $L{\ddot{u}}beck$, Germany). Results: A total of 412 HCW participated in the study. Among them, 14 were excluded due to the inadequate sample amount or medical history not secured. Of the 398 HCWs analyzed, 16.6% (66/398) were men and the mean age was $33.82{\pm}9.10years$ (range, 21-67). The mean anti-PT IgG titer was $8.32{\pm}20.40IU/mL$ (range, 0.4-287.5 IU/mL). The overall seroprevalence (rate of anti-PT IgG antibody [Ab] titer > 5 IU/mL) was 33.7%. Three (0.8%) HCWs had the Ab level > 100 IU/mL indicated acute infection. There was no statistically significant difference in the seroprevalence and mean titer of anti-PT IgG Ab according to age group, type of occupation, patient-facing position, or working in the pediatric department. Conclusion: The seroprevalence of pertussis of the HCWs of a university hospital in Korea was low, and there were some unrecognized acute infections. Therefore, booster immunization of pertussis to HCWs should be actively considered.

Synthesis of New 4-Oxo-2-Thioxo-1,2,3,4-Tetrahydropyrimidine Derivatives with an Incorporated Thiazolidinone Moiety and Testing Their Possible Serine Protease and Cercarial Elastase Inhibitory Effects with a Possible Prospective to Block Penetration of Schistosoma mansoni Cercariae into the Mice Skin

  • Bahgat Mahmoud Mohamed;Maghraby Amany Sayed;Heiba Mogeda Emam;Ruppel Andreas;Fathalla Omar Abd-elfattah Mohamed
    • Archives of Pharmacal Research
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    • v.28 no.9
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    • pp.1002-1012
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    • 2005
  • 5-Substituted 4-oxo-2-thioxo-1,,2,3,4-tetrahydropyrimidine were synthesized by interaction of 4­oxo-2-thioxo-1,2,3,4-tetrahydropyrimidine-5-sulfonylhydrazide with some aldehydes to give the corresponding Schiff-bases, which after cyclization gave corresponding thiazolidinones. For some of the thiazolidinones, Mannich bases reaction was carried out. All the derivatives were tested for their possible inhibitory effect on Schistosoma mansoni cercarial elastase (CE). Only, N'-(4-methylbenzyledine)-4-oxo-2-thioxo-1,2 ,3,4-tetrahydropyrimidine-5-sulfonylhydrazide was found to have potent inhibitory effect on the CE activity with $IC_{50} = 264{\mu}M.$ Upon its use as a paint for mice tails before infection with S. mansoni cercariae, the compound formulated in jojoba oil caused a significant reduction ($93\%$; P-value = 0.0002) in the worm burden. IgG & IgM in mice sera were measured by using several S. mansoni antigens by ELISA. Sera from treated infected mice (TIM) 2, 4, and 6 weeks (W) post infection (PI) showed 1.2 folds lower, 1.2 folds higher, 1.7 folds lower IgM reactivity against soluble cercarial antigenic preparation (CAP), respectively, when compared with sera collected from infected untreated mice (IUM). Sera from TIM 2, 4, and 6WPI showed 1.3, 1.6, and 1.7 folds higher IgG reactivity, respectively against CAP than the IgG reactivity from IUM. Sera from TIM 2, 4 and 6WPI showed 1.5, 1.2 folds lower and 1.4 folds higher IgM reactivity, respectively against soluble worm antigenic preparation (SWAP) when compared with sera collected from IUM. Sera from TIM 2, 4, and 6WPI showed 1.4, 1 folds lower and 1 fold higher IgG reactivity, respectivley to SWAP when compared with sera from IUM. Sera from TIM 2, 4, and 6WPI had generaly lower IgM and IgG reactivities against soluble egg antigen (SEA) when compared with sera from IUM.

Productivity, Isolation and Purification of Egg Yolk Antibody(IgY) againt Food Poisoning Bacteria (Salmonella typhimurium) (식중독균 항원(Salmonella typhimurium)에 의한 계란항체(IgY) 생산성과 분리 정제)

  • 한준표;백반석;배만종
    • Journal of the East Asian Society of Dietary Life
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    • v.9 no.2
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    • pp.200-206
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    • 1999
  • This study was carried out to get a industrial information about a possibility of IgY antibody production, antimicrobial activity and Properties of IgY antibody in egg yolk. After the initial immunization the anti-Salmonella typhimurium IgY antibody level gradually were decreased from firth week to tenth week. On the other hand, the antibody level in the serum were increased from the first week, reaching its peak in the sixth week. Molecular weights of IgY were estimated approximately 72-75KD in a heavy chain and 30-40KD in a light chain by electrophoresis.

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Determination of Ovalbumin in Processed Foods by Immunological Methods

  • Seo, Ji-Hyun;Lee, Ju-Woon;Kang, Sin-Bok;Lee, Ha-Baik;Yook, Hong-Sun;Kim, Mee-Ree;Kim, Kee-Hyuk;Byun, Myung-Woo
    • Preventive Nutrition and Food Science
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    • v.7 no.4
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    • pp.373-377
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    • 2002
  • Allergens in processed foods may place persons with food allergies at significant risk when the labels do not Provide sufficient warnings or identification of high-risk ingredients. Because egg proteins are common food allergens, this study was carried out to identify hen's egg albumin (ovalbumin, OVA) in five commercially processed foods containing egg (custayd, cookie and pasta), and chicken meat (sausage and meatball) by immunological methods using commercially produced murine monoclonal immunoglobulin G (M-IgG), immunoblotting and enzyme linked immunosorbent assay (ELISA). Sample buffer with chelating and reducing agents was prepared and used for the preparation of the protein fractions from the foods. Most bands in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) profile (5~15% gradient gel) presented at 75 kDa below. OVA (43 kDa) in the sample lanes could not be visually observed on the gel. However, OVA in solutions prepared from custard and cookie could be detected by M-IgG, but were not detected in sausage and pasta. OVA in all samples could be quantitatively determined by the equation obtained from the standard curve by ELISA. Cookie and custard containing egg white and egg, respectively, contained very high concentrations of OVA. OVA in the other products were present in relatively low concentrations, but sufficiently high to pose possible risk of allergy, ELISA is a very sensitive and precise method for the identification and quantification of allergens in food products including allergy-inducible materials.

DA-9601, Artemisia Asiatica Herbal Extract, Ameliorates Airway Inflammation of Allergic Asthma in Mice

  • Kim, Ji Young;Kim, Dae Yong;Lee, Yun Song;Lee, Bong Ki;Lee, Kyung-Hoon;Ro, Jai Youl
    • Molecules and Cells
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    • v.22 no.1
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    • pp.104-112
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    • 2006
  • We previously reported that DA-9601, ethanol herbal extract of Artemisia asiatica, inhibited histamine and leukotriene releases in guinea pig lung mast cells activated with specific antigen/antibody reaction. This study aimed to evaluate the inhibitory effect of DA-9601 on the OVA-induced airway inflammation in allergic asthma mouse model. BALB/c mice were sensitized and challenged with OVA. DA-9601 was administered orally 1 h before every local OVA-challenge. OVA-specific serum IgE was measured by ELISA, recruitment of inflammatory cells in BAL fluids and lung tissues by Diff-Quik and H&E staining, respectively, the expressions of CD40, CD40L and VCAM-1 by immunohistochemistry, goblet cell hyperplasia by PAS staining, activities of MMPs by gelatin zymography, expressions of mRNA and proteins of cytokines by RT-PCR and ELISA, activities of MAP kinases by western blot, and activity of NF-${\kappa}B$ by EMSA. DA-9601 reduced IgE level, recruitment of inflammatory cells into the BAL fluid and lung tissues, expressions of CD40, CD40L and VCAM-1 molecules, goblet cell hyperplasia, MMPs activity, expressions of mRNA and productions of various cytokines, activities of MAP kinases and NK-${\kappa}B$ increased from OVA-challenged mice. These data suggest that DA-9601 may be developed as a clinical therapeutic agent in allergic diseases due to suppressing the airway allergic inflammation via regulation of various cellular molecules expressed by MAP kinases/NF-${\kappa}B$ pathway.

Variations of Immunoglobulins in Colostrum and Immune Milk as Affected by Antigen Releasing Devices

  • Zhaoa, Shengguo;Zhanga, Chungang;Wang, Jiaqi;Liu, Guanglei;Bu, Dengpan;Cheng, Jinbo;Zhou, Lingyun
    • Asian-Australasian Journal of Animal Sciences
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    • v.23 no.9
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    • pp.1184-1189
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    • 2010
  • This work was conducted to examine the variation of immunoglobulins (Igs) in serum, immune milk, normal milk and colostrum upon implantation of a new Antigen Releasing Device (ARD). The core of each ARD housed an immunostimulating complex (ISCOM) that was made of adjuvant Quil A and type XIII lipase from a Pseudomonas sp. Each ARD was coated with polylactic acid, known as polylactide, that controls antigen release. Twenty lactating Chinese Holstein cows were divided into 2 groups (n = 10): test group and control group. All cows in the test group were implanted with a single injection in the right iliac lymph node with 3 types of ARDs, which were designed to release the antigens at d 0, 14 and 28 post-implantation. Blood and milk samples were collected from both groups, and colostrum samples were also collected from other post-partum cows in the same farm. Concentrations of $IgG_1$, IgA and IgM in whey and serum were measured by sandwich ELISA. The results showed that the $IgG_1$, IgA and IgM concentrations in serum and whey from the test group were higher than from the control group. Among the three Igs measured, the $IgG_1$ concentration in serum was significantly higher at d 40 after ARD implantation, and the $IgG_1$ concentration in whey peaked at d 9, 17 and 30, which corresponded with release of the antigen. Based on Pearson's correlation between Ig concentration and production parameters, IgA concentration in normal milk was positively correlated with lactation period, which reflected IgA changes during the lactation period in immune milk. In colostrum, $IgG_1$, IgA and IgM decreased abruptly from d 0 to 3, and then decreased slightly. In conclusion, serum $IgG_1$ concentration can be affected by controlled release of the ARD, while whey IgA levels are primarily affected by lactation period. These results may be useful in future studies designed to regulate concentrations of Igs in immune milk.

Generation of polyclonal antiserum to olive flounder (Paralichthys olivaceus) immunoglobulin by immunization of rabbit with plasmids containing heavy chain gene of olive flounder immunoglobulin

  • Kim, Gi-Hong;Kwon, Se-Ryun;Kim, Chun-Soo;Lee, Eun-Hye
    • Journal of fish pathology
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    • v.19 no.2
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    • pp.183-188
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    • 2006
  • In fish vaccinology, the secondary antibodies against fish immunoglobulins (Igs) are necessary to measure specific humoral immune responses in immunized fish. In the present study, polyclonal antiserum against olive flounder (Paralichthys olivaceus) IgM heavy chain was generated by intramuscular immunization of rabbit with Escherichia coli/eukaryotic shuttle vector containing open reading frame (ORF) of olive flounder IgM heavy chain. Western blot analysis demonstrated the specific activity of the rabbit antiserum with reduced olive flounder serum H chain at dilutions up to 1:1000. Titer of immunized rabbit serum against olive flounder serum was significantly higher than that of pre-immunized rabbit serum when determined by ELISA.