• Title/Summary/Keyword: enzyme-linked immunosorbent assay(ELISA)

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Prevalence of bovine viral diarrhea virus from Korean native cattle farms in Jeju (제주지역 한우의 소 바이러스성 설사병 바이러스 감염실태)

  • Seong-Cheol Cho;Hyoung-Seok Yang;Changnam Park;Si-Taek Kim;Eun-Ju Ko;Won-Geun Son
    • Korean Journal of Veterinary Research
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    • v.63 no.2
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    • pp.12.1-12.7
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    • 2023
  • Bovine viral diarrhea virus (BVDV) is an RNA virus belonging to Pestivirus in the family Flaviviridae. BVDV has economic significance for the livestock industry because of its association with acute disease, fetal loss, and birth of persistently infected (PI) animals. This study aimed to investigate the BVDV infection rates in Korean native cattle farms in Jeju for further planning of a BVDV control program in the Jeju Province. BVDV antibodies and antigens were tested in 15,842 sera collected from 302 Korean native cattle herds between January 2014 and June 2017 using enzyme-linked immunosorbent assay (ELISA). Viral antigen was detected by reverse transcription-polymerase chain reaction from 60 sera that were antigen ELISA-positive. BVDV antibodies were found in 90.7% (274/302) herds and 61.1% (9,678/15,842) cows. BVDV antigens were found in 13.2% (40/302) herds and 0.4% (61/15,842) cows. The oldest animal group (> 8 years) exhibited the highest sero-positive rates (91%), while the youngest animal group (< 1 years) had the highest antigen positivity rates (0.52%). Of the 60 antigen-positive sera, BVDV types 1 and 2 were found in 36 and 12 sera, respectively. Additionally, six animals were considered to be PI as BVDV was continually detected in annual examination.

Serological Studies on the Specific Antibodies Against P-pili of Uropathogenic Escherichia coli (요로 감염환자에서 혈청학적 방법을 이용한 P-pili특이혈중 항체의 조사)

  • 이원용;김종배
    • Biomedical Science Letters
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    • v.2 no.1
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    • pp.31-40
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    • 1996
  • Escherichia coli is one of the most common etiological agents in urinary tract infection. An important virulence factor is the adhesive capacity of E. coli to uroepithelial cell, mediated by bacterial fimbriae. The Adhesion property has been regarded as an important virulence determinant in urinary tract infections. A total of 60 patients, who were diagnosed microbiologically as urinary tract infections, were examined by immunoblotting and enzyme-linked immunosorbent assay(ELISA). Uropathogenic E. coli with recombinant plasmid were positive for mannose resistant hemagglutination (MRHA). For identification of p-fimbriae subtype in uropathogenic E. coli, In the immunoblot analysis, specific bands in the range of p-fimbriae molecular weight of 17KD-22KD were identified. For the distribution of p-fimbriae subtype in the patient sera, 34/60(56.7%) were positive for $F7_1$, 28/60(46.7%) were positive for $F7_2$, and 30/60(50%) were positive for F13 with immunoblotting method. similar trends were observed in the enzyme-linked immunosorbent assay. Relatively good specificity(92.6%) and sensitivity(90%) were found in the ELISA test system using mixed antigens of purified $F7_1$, $F7_2$, and F13 p-fimbriae, and 60 sera from patients with urinary tract infections. In conclusion The serological tests were convenient method in diagnosis of urinary tract infections. among those ELISA could be recommended in diagnosis of urinary tract infections.

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Development of Immunoassay Systems for the Assay of Soy Protein in Meat Products; The Assay of Soy Protein in Meat Blends and Commercial Product by Enzyme-linked Immunosorbent Assay(ELISA) (육제품에 첨가된 대두단백 정량을 위한 면역분석법 개발에 관한 연구 ; ELISA에 의한 고기유화물 및 시판육제품에 첨가된 대두단백 정량)

  • Kim, Cheon-Jei;Kim, Jong-Bae;Kim, Byung-Cheol;Lee, Seoung-Bae;Jung, Sung-Won;Choe, Doo-Young;Ko, Won-Sick
    • Korean Journal of Food Science and Technology
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    • v.24 no.5
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    • pp.397-403
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    • 1992
  • This study was carried out for the development of assay method to quantify the soy protein content in meat homogenate, emulsion-type sausage and commercial meat products by Enzyme-linked Immunosorbent Assay(ELISA). The standard antigen was extracted before and after heat treatment. It was observed that the degree of reaction was not varied significantly according to the heating temperature. The recovery rate in meat homogenate and emulsion-type sausage was not varied significantly according to the heating temperature. The reaction was not interfered with fat and spices of the samples. Samples with 10% soy protein showed lower correlation than those with 2% and 5% soy protein. The recovery rate in commercial meat products showed difference individually. The correlation of some products with raw vegetable and wheat starch was relatively low.

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Application of the Enzyme-linked Immunosorbent Assay to the Serodiagnosis of Typhoid Fever (장티푸스의 혈청학적 진단에 효소결합면역측정법(Enzyme-linked Immunosorbent Assay)의 적용 실험)

  • Kye, Ki-Shik;Kim, Yae-Hum;Choi, Kang-Won;Hwang, Eung-Soo;Kook, Yoon-Ho;Lee, Seung-Hoon;Cha, Chang-Yong
    • The Journal of the Korean Society for Microbiology
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    • v.18 no.1
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    • pp.73-85
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    • 1983
  • The advantages of the enzyme-linked immunosorbent assay(ELISA) are its sensitivity and its simplicity in detecting IgM and IgG antibodies. For applying the ELISA to the diagnosis of typhoid fever, first of all, experiments were performed to determine which concentration of killed whole cell antigens and lipopolysaccharide(LPS) antigens of S. typhi(0.901 w) were optimally coated to the wells of the polystyrene and polyvinylchloride microplate, using the hyperimmune sera from rabbits against S. typhi. By using both kinds of antigens of S. typhi adsorbed to the ELISA microplate, the changing patterns of IgG and IgM antibodies in the sera from rabbits responding to the killed whole cell antigens of S. typhi(0901 w) during the prolonged immunization were serially traced by the ELISA. At the same time, the level of antibodies against S. typhi in sera fron patients with typhoid fever and from normal healthy persons were measured by the ELISA employing the killed whole cell antigens and LPS antigens as the coating antigens. The results obtained were summerized as follow: 1. The optimal concentration of the killed whole cell antigens, which were more easily adsorbed to the polystyrene plate than the polyvinylchloride plate, was $10^8cells/ml$ of carbonate buffer(pH. 9.6) on the wells of the polystyrene plate when treated at $37^{\circ}C$ for 4 hours. On the other hand, the optimal concentration of lipopolysaccharide antigens, which were adsorbed only to the polyvinylchloride plate, was $100{\mu}g/ml$ of carbonate buffer(pH. 9.6) on the wells of the polyvinylchloride plate when treated at $37^{\circ}C$ for 4 hours. 2. IgM antibody response were dominating in rabbits responding to the killed whole cell antigens of S. typhi(0.901 w), and were more specific to the LPS antigens than to the killed whole cell antigens in the ELISA. Good correlations were made between the IgM titers by the ELISA and the aggglutinating titers of sera from the immunized rabbits. 3. Both IgG and IgM agglutination titers by the ELISA in sera from most of patients with typhoid fever were above 1:320 but those in sera from most of normal, healthy persons were below 1:80. 4. There were close correlations between the antibody titers by the ELISA and the agglutinating titers to the killed whole cell antigens in the tested human sera, IgM titers being more correlated with the agglutinating titers than IgG titers. But a little correlations were made between the antibody titers by the ELISA and the agglutinating titers to the LPS antigens. 5. IgM titers in the tested human sera were similar to IgG titers detected by the ELISA employing the killd whole cells antigens and the LPS antigens. 6. Good correlations were made between the antibody titers demonstrated by the ELISA performed on the killed whole cell antigens and the LPS antigens as the different, coating antigens on the ELISA microplates.

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Comparative Diagnostic Studies on Serologic and Molecular Biological Tests Against Haemorrhagic Fever with Renal Syndrome (신증후출혈열 환자의 혈청학적 및 분자생물학적 진단 검사법 비교)

  • 우영대;문희주;배형준
    • Biomedical Science Letters
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    • v.6 no.2
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    • pp.141-149
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    • 2000
  • The etiologic agents of haemorrhagic fever with renal syndrome (HFRS) in Korea are Hantaan and Seoul virus in the genus Hantavirus, family Bunyaviridae. Antibody titers of sera from HFRS patients against Hantaan virus were measured by immunofluorescent antibody technique (IFAT), enzyme-linked immunosorbent assay (ELISA), high density composite particle agglutination (HDPA) and plaque reduction neutralization test (PRNI). PRNT and nested reverse transcriptase polymerase chain reaction (nested RT-PCR) was used for serotypic differentiation of Hantaviruses against Hantaan and Seoul virus. Eight doubtful HFRS patients showed higher fluorescent, IgG ELISA, agglutination and neutralizing antibody titer by IFAT, ELISA IgG, HDPA and PRNT, respectively Five out of them showed high IgM antibody titer by IgM capture ELISA against Hantaan virus, remarkably. Fifteen HFRS patients showed higher fluorescent antibody titer by IFAT. In PRNT, 12 out of them showed high neutralizing antibody titer against HTNV, 2 against SEOV and 1 against both viruses. In nested RT-PCR using serotype specific-primer, 3 out of them showed positive against HTNV and 1 against SEOV.

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

  • 최석호
    • Food Science of Animal Resources
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    • v.24 no.2
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    • pp.176-181
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    • 2004
  • The specificity of sandwich enzyme-linked immunosorbent assay (ELISA) to detect Salmonella in milk was determined in this study. The antibodies used in sandwich ELISA were egg yolk immunoglobulin G (IgY) obtained after immunization of hen with outer membrane protein (OMP) fraction from Salmonella typhimurium and rabbit IgG obtained after immunization of rabbit with the purified OMP with the molecular weight of 40,000. The immunoblot assay showed that the IgY reacted strongly with OMP with the molecular weight of 6,000 and the rabbit IgG reacted strongly with OMP with the molecular weights of 40,000, 35,000, and 6,000 from the bacteria including Salmonella which belongs to Enterobacteriaceae. The IgY and rabbit IgG also reacted with other proteins from Salmonella typhimurium in immunoblot assay. Competitive ELISA showed that IgY showed specifity to react with two strains of Salmonella typhimurium and Salmonella cholerasuis but not with Escherichia coli and Yersinia enterocolitica. Two strains of Salmonella typhimurium added to UHT milk showed the highest absorbance of all the bacteria used in the sandwich ELISA. Some strains of Salmonella cholerasuis showed higher absorbances than non-Salmonella bacteria.

Diagnosis of Potato Leafroll disease by Fluorescence Microscopic Detection of Callose Stained with Resorcin Blue (Resorcin Blue 염색 기법에 의한 감자 잎말림병의 형광 현미경적 진단)

  • 이철호;나용준
    • Korean Journal Plant Pathology
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    • v.11 no.2
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    • pp.101-106
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    • 1995
  • Deep blue fluorescence of resorcin blue-stained callose was observed only in the potato leafroll virus (PLRV)-infected potato plants, but not in other potato viruses investigated. The plant sections stained with aniline blue showed non-specific fluorescence regardless of PLRV infection, which means that aniline blue is not suitable for the staining of callose induces by PLRV infection. The fluorescence of resorcin blue-stained callose was more easily detectable than autofluorescence by a direct fluorescence detection method because of its deep blue color. The lateral branch of lower leaves was turned out to be the best material for fluorescence observation of all plant parts tested. In comparison of diagnostic efficacy of this technique to enzyme-linked immunosorbent assay (ELISA), PLRV infected potato plants showed corresponding increment of the fluorescence of resorcin blue stained callose as absorption values in ELISA increased. In the future, the criteria measuring the fluorescence objectively are thought to be determined for the practical application to the diagnosis of potato leafroll disease.

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A serological survey on large outbreak of bovine brucellosis in dairy farm (부루셀라병 다두이환 목장의 혈청항체가 조사)

  • 김상윤;김정화;김대원
    • Korean Journal of Veterinary Service
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    • v.22 no.2
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    • pp.159-167
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    • 1999
  • This survey was conducted for the serological confirmation on large outbreak of bovine brucellosis in two dairy farms. Serological tests were performed by the plate agglutination test, tube agglutination test, enzyme linked immunosorbent assay(ELISA), complement fixation, ,test(CFT) and rose bengal plate test(RBPT). Total 200 heads(134 heads in farm A and 66 heads in farm B) were tested. The primigravida and positive group have been raised separately in the farm A and both group have been raised together in the farm B.. The result were summarized as follows ; 1. Positive ratios in positive herds of farms by the tube agglutination test were 68.3% in farm A and 53.2% in farm B. 2. Seroconversion to brucella was observed in the primigravidas group in farm B, but was not observed in the primigravidas group in farm A. 3. All calves born in positive herd were serologically negative at time of test. 4. Positive ratio of ELISA in farm A was higher than that of tube agglutination test. 5. Number of positive reactors by the CFT, RBPT in farm A were equal to those of tube agglutination test.

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Survey on the antibody distributions of swine pneumonia by ELISA in Daegu province (효소면역흡착법을 이용한 대구지역의 돼지폐렴에 대한 항체분포조사 연구)

  • 조유정;서동균;송동준;이춘식;배영찬
    • Korean Journal of Veterinary Service
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    • v.25 no.3
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    • pp.237-243
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    • 2002
  • The study was performed to investigate the antibody distributions of 4 swine respiratory disease including M hyopneumoniae, P multocida, A pleuropneumoniae serotype 2 and 5 in Daegu area by enzyme-linked immunosorbent assay(ELISA). 1. The overall sero-positive rates were 55.6% in June, 48.0% in August, 51.3% in October and 25.4% in November. 2. The positive reaction rates to M hyopneumoniae, P multocida, A pleuropneumoniae serotype 2 and 5 were found to be 50.0%, 36.5%, 55.0%, and 42.0% respectively. 3. The antibody titers were distributed on range 20~80 in M hyopneumoniae, 20~80 in P multocida, 160~640, 20~80 in A pleuropneumoniae serotype 2 and 5.

Production of Group Specific Monoclonal Antibody to Aflatoxins and its Application to Enzyme-linked Immunosorbent Assay

  • Kim, Sung-Hee;Cha, Sang-Ho;Karyn, Bischoff;Park, Sung-Won;Son, Seong-Wan;Kang, Hwan-Goo
    • Toxicological Research
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    • v.27 no.2
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    • pp.125-131
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    • 2011
  • Through the present study, we produced a monoclonal antibody against aflatoxin B1 (AFB1) using AFB1-carboxymethoxylamine BSA conjugates. One clone showing high binding ability was selected and it was applied to develop a direct competitive ELISA system. The epitope densities of AFB1-CMO against BSA and KLH were about 1 : 6 and 1 : 545, respectively. The monoclonal antibody (mAb) from cloned hybridoma cell was the IgG1 subclass with ${\lambda}$-type light chains. The $IC_{50}s$ of the monoclonal antibody developed for AFB1, AFB2, AFG1 and AFG2 were 4.36, 7.22, 6.61 and 29.41 ng/ml, respectively, based on the AFB1-KLH coated ELISA system and 15.28, 26.62, 32.75 and 56.67 ng/ml, respectively, based on the mAb coated ELISA. Cross-relativities of mAb to AFB1 for AFB2, AFG1 and AFG2 were 60.47, 65.97 and 14.83% in the AFB1-KLH coated ELISA, and 59.41, 46.66 and 26.97% in the mAb coated ELISA, respectively. Quantitative calculations for AFB1 from the AFB1-Ab ELISA and AFB1-Ag ELISA ranged from 0.25 to 25 ng/ml ($R^2$ > 0.99) and from 1 to 100 ng/ml ($R^2$ > 0.99), respectively. The intra- and inter-assay precision CVs were < 10% in both ELISA assay, representing good reproducibility of developed assay. Recoveries ranged from 79.18 to 91.27%, CVs ranged from 3.21 to 7.97% after spiking AFB1 at concentrations ranging from 5 to 50 ng/ml and following by extraction with 70% methanol solution in the Ab-coated ELISA. In conclusion, we produced a group specific mAb against aflatoxins and developed two direct competitive ELISAs for the detection of AFB1 in feeds based on a monoclonal antibody developed.