• Title/Summary/Keyword: indirect enzyme-linked immunosorbent assay

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Serological Study on the Cross-Reactivity of Bacteroides gingivalis, Bacteroides intermedius and Bacteroides asaccharolyticus by Indirect Immunofluorescence and Enzyme- Linked Immunosorbent Assay (형광 현미경법 및 효소결합 면역흡착법을 이용한 Bacteroides gingivalis, Bacteroides intermedius 및 Bacteriudes asaccharolyticus의 혈청학적 연구)

  • Chung, C.P.;Lee, J.Y.;Lee, Y.H.;Chung, H.W.;Chung, H.J.
    • The Journal of the Korean Society for Microbiology
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    • v.22 no.2
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    • pp.117-123
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    • 1987
  • Previous studies have been performed for the sero-identification of selected species of Bacteroides by immunofluorescence antibody techniques and enzyme-linked immunosorbent assay using species-specific rabbit antisera to B. gingivalis, B. intermedius, and B. melaninogenicus. However, these studies have not commented on the serological cross-reactivity between these 3 species of black- pigmented Bacteroides. For the cross-reactivity study, antisera to B. gingivalis ATCC 33277, B. intermedius ATC C25261 and B. asaccharolyticus ATCC 25260 were raised from rabbits. Preliminary study for observing the cross-reactivity between these species was performed by indirect immunoflourescence technique. Immunoabsorption of the antisera was done with bacterial cells from the other species and the species-specificity of the antisera was conformed by the absence of reactivity with bacterial strains from the other species by indirect immunofluorescence technique and enzyme-linked immunosorbent assay. Three representative unabsorbed antisera cross-reacted strongly with cells from the other species. Especially, anti-B. asaccharolyticus ATCC 25260 antiserum showed a strong cross-reactivity with B. gingivalis ATCC33277. After immunoabsorption of 3 representative antisera with the other species, the cross-reactivity was found only between B. gingivalis ATCC 33277 and B. asaccharolyticus ATCC 25260. Further study would be necessary to clarify the cross-reactivity between important oral black-pigmented Bacteroides from subgingival plaque or bacterial colonies for rapid identification.

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Quantification of Reproductive Output of the Butter Clam, Saxidomus purpuratus(Sowerby, 1852) Using Enzyme-Linked Immunosorbent Assay (ELISA)

  • Park, Kyung-Il;Choi, Jin-Woo;Choi, Kwang-Sik
    • Ocean and Polar Research
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    • v.25 no.3
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    • pp.249-256
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    • 2003
  • An immunological method was developed in this study to quantify reproductive output of the female butter clam, Saxidomus purpuratus. A clam egg-specific polyclonal antibody was developed using the purified butter clam egg as an antigen. An indirect Enzyme-Linked Immunosorbent Assay (ELISA) was used in quantitative measurement of the eggs. Size of the butter clam eggs ranged from $70.81{\pm}7.52{\mu}m$ in histology or $88.56{\pm}11.31{\mu}m$ in intact eggs. The predominant egg constituent was protein (37.44%), followed by lipids (11.40%) and carbohydrates (9.68%). The SDS-PAGE showed that the egg proteins are composed of several peptides of molecular weights consisting of 247, 200, 99, 91, 54 and 47 kDa. ELISA indicated that the clams collected from Geoje Island in May 2002 produced 8.2 to 26.8% of their body weight as eggs or 9,307,309 to 31,156,333 with a mean of 16,931,893 eggs per individual clam. The results of this study thus suggest that indirect ELISA using rabbit anti-clam egg IgG as a primary antibody is a rapid, affordable and sensitive method to assess reproductive output of 5. purpuratus and possibly other bivalves using a small amount of eggs.

Quantitative Analysis of Phosphinothricin-N-acetyltransferase in Genetically Modified Herbicide Tolerant Pepper by an Enzyme-Linked Immunosorbent Assay

  • Shim, Youn-Young;Shin, Weon-Sun;Moon, Gi-Seong;Kim, Kyung-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.17 no.4
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    • pp.681-684
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    • 2007
  • An immunoassay method was developed to quantitatively detect phosphinothricin-N-acetyltransferase (PAT) encoded by the Bialaphos resistance (bar) gene in genetically modified (GM) pepper. The histidine-tagged PAT was overexpressed in Escherichia coli M15 (pQE3l-bar) and efficiently purified by $Ni^{2+}$ affinity chromatography. A developed sandwich enzyme-linked immunosorbent assay (S-ELISA) method (detection limit: $0.01{\mu}g/ml$) was 100-fold more sensitive than a competitive indirect ELISA (CI-ELISA) method or Western blot analysis in detecting the recombinant PAT. In real sample tests, PAT in genetically modified herbicide-tolerant (GMHT) peppers was successfully quantified [$4.9{\pm}0.4{\mu}g/g$ of sample (n=6)] by the S-ELISA method. The S-ELISA method developed here could be applied to other GMHT crops and vegetables producing PAT.

Production and Characterization of Monoclonal Antibodies to a Generic Hapten for-Class-Specific Determination of Organophosphorus Pesticides

  • Jang, Mi-Soon;Lee, Soo-Jung;Xue, Xiaoping;Kwon, Hyuk-Man;Ra, Choon-Sup;Lee, Yong-Tae;Chung, Tae-Wan
    • Bulletin of the Korean Chemical Society
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    • v.23 no.8
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    • pp.1116-1119
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    • 2002
  • Monoclonal antibodies have been generated against a generic hapten, ο,ο-diethyl ο-(5-carboxy-2-fluorophenyl) phosphorothioate, for the determination of organophosphorus (OP) pesticides in a class-specific manner. In an indirect competitive enzyme-linked immunosorbent assay (ELISA) format, employing a heterologous coating antigen, these monoclonal antibodies showed desirable properties for use in the class-specific determination, i.e., broad specificity and high sensitivity. The IC50 values of four commonly used ο,ο-diethyl OP pesticides were fairly uniform ranging from 0.1 to 0.3 ㎛/mL. The IC50 values of three ο,ο-dimethyl derivatives were between 0.3 and 1.4 ㎛/mL. These values, together with the limits of detection (LOD), were better, in terms of the specificity and sensitivity, compared with the values obtained previously with polyclonal antibodies.

An Enzyme-Linked Immunosorbent Assay for Detection of Pantothenic Acid (판토텐산의 분석을 위한 효소면역측정법)

  • Shon, Dong-Hwa;Park, Youn-Sick;Bae, Gun-Won
    • Korean Journal of Food Science and Technology
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    • v.32 no.5
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    • pp.1009-1014
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    • 2000
  • In order to detect pantothenic acid (PA), conditions for enzyme-linked immunosorbent assay (ELISA) were established. Anti-PA-BSA antibody was produced from rabbits immunized with PA-bovine serum albumin (BSA) conjugates which were prepared by the bromoacetyl chloride [Bc] method (PA-BSA[Bc]) and by the periodate oxidation [Po] method (PA-BSA[Po]). PA-BSA[Bc] and PA-BSA[Po] was used as a coating antigen for competitive indirect(ci)ELISA. The Anti-PA-BSA[Po] antibody on ciELISA showed no competitive reaction. The detection limit of PA by ciELISA using Anti-PA-BSA[Bc] antibody was 1 ppm. The Anti-PA-BSA[Bc] antibody showed little cross-reactivity to PA derivatives such as pantoyllactone, pantetheine, pantothenyl alcohol, and acetyl CoA. The detection limit of PA by microbiological assay (MBA) was 10 ppb. Assay recoveries of PA in egg, cow's liver, and lettuce by ciELISA were 109, 64, and 344%, respectively, comparing with the MBA results.

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Serological prevalence of brucellosis of cattle in selected dairy farms in Bangladesh

  • Hassan, Abdullah Al;Uddin, M. Bashir;Islam, Md. Rafiqul;Cho, Ho-Seong;Hossain, Md. Mukter
    • Korean Journal of Veterinary Research
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    • v.54 no.4
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    • pp.239-243
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    • 2014
  • This study was conducted to investigate the status of brucellosis in dairy cattle from five selected dairy farms in the Mohammadpur Beribadh area of Bangladesh. A cross-sectional study was carried out from October 2010 to March 2011 in which a total of 334 serum samples from cattle in five herds were screened by the Rose-Bengal plate-agglutination test (RBPT) and the positives were confirmed using an indirect enzyme-linked immunosorbent assay (I-ELISA). A structured questionnaire was used to collect epidemiological information describing the animals. Overall, 4.20% of the animals were RBPT positive, while subsequent confirmatory tests with I-ELISA revealed that the overall animal-level prevalence derived from the samples was 1.20%. Additionally, the prevalence was relatively higher in females than in males. A significant association was found between abortion, age of the animals, and the occurrence of brucellosis (p < 0.05). Considering the overall low prevalence of brucellosis in the selected farms in the present study, a brucellosis eradication program for dairy farms using a test-and-slaughter policy would be possible.

Detection of PCB77 by Indirect Competitive Enzyme-linked Immunosorbent Assay in Sea Sediment Samples

  • Chen, Han-Yu;Zhuang, Hui-Sheng;Yang, Guang-Xin
    • Bulletin of the Korean Chemical Society
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    • v.34 no.3
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    • pp.922-926
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    • 2013
  • 3,3',4,4'-Tetrachlorobiphenyl (IUPAC PCB77) is one of seven indicative polychlorinated biphenyls (PCBs) in the surface sediments. The current study presents a novel polyclonal antibody for the determination of the PCB77 using indirect competitive enzyme-linked immunosorbent assay. Under optimum conditions, PCB77 was determined within the concentration range of 0.01-100 ${\mu}g\;L^{-1}$, with a detection limit of 0.057 ${\mu}g\;L^{-1}$. The assays were tested for their cross-reactivity profiles using 3 selected congeners and 4 Aroclor products. The assays were highly specific for coplanar PCB congeners, but less specific for Aroclor1248. The spiked recoveries from five sediment samples were 86%-114% for PCB77 from ELISA, which were satisfactory. The current study demonstrated that the developed antiserum and immunoassay procedure can be used to detect PCB77 in environmental samples. The results of the sediment analysis were confirmed by conventional GC/ECD.

Detection of antibodies against infectious Borna disease virus -a comparison of three serological methods- (보르나병 바이러스 항체검출을 위한 연구 -세 가지 혈청진단법의 비교-)

  • Lee, Du-sik
    • Korean Journal of Veterinary Research
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    • v.32 no.1
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    • pp.57-61
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    • 1992
  • To determin the accuracy of serological methods in detecting Borna-disease(BD) viral antibodies, 273 experimentally infected rabbit sera were compared by using indirect immunofluorescence antibody test(IFA), serum neutralization test(SN) and enzyme-linked immunosorbent assay(ELISA). One hundred twenty-three serum samples had BD viral antibodies detected by IFA. CELISA antibodies to BD virus were also present in the same one hundred twenty-three serum samples. However, neutralization test antibodies to BD virus were present in 27 of the in rabbit serum samples. Neutralization test was sensitive in comparison with KFA and CELISA. In comparison with IFA, CELISA was both sensitive and specific in detecting BD viral antibodies. These results extend observations made with laboratory animals to the diagnosis of naturally infected animals.

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Indirect enzyme linked immunosorbent assay for the diagnosis of brucellosis in cattle

  • Rahman, Siddiqur;Huque, Fazlul;Ahasan, Shamim;Song, Hee-Jong
    • Korean Journal of Veterinary Service
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    • v.33 no.2
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    • pp.113-119
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    • 2010
  • Brucellosis is a major zoonosis caused by Gram negative facultative intracellular bacterial organisms of the genus Brucella that are pathogenic for a wide variety of animals and human beings. Because of its economic impact on animal health and the risk to the human population,most countries have a brucellosis control program. Brucellosis is also an economically important andprevalent disease in Bangladesh. The accurate and prompt diagnosis is very important in controlling and eradicating of the disease in animals. The present study was undertaken to determine the seroprevalence of brucellosis in cattle in Mymensingh and Patuakhali district of Bangladesh. A total of 120 serum samples were collected from the two districts along with a questionnaire related to the epidemiology of the disease. The sampleswere screened by using slow agglutination test and conformed by indirect enzyme linked immunosorbent assay. The overall seroprevalence of brucellosis in cattle was 5% and it was observed that, a higher prevalence of Brucella was found in female than male, through natural breeding than artificial insemination (AI) and animal above 4 years old are highly susceptible than younger ones. Higher prevalence was found in aborted animals in comparison with non aborted animal. Finally, the study revealed that the female animal has more susceptible to brucellosis and healthy semen should be used for AI.

Oral Manifestation of Paraneoplastic Pemphigus

  • Kim, Seurin;Park, In Hee;Park, YounJung;Kwon, Jeong-Seung;Choi, Jong-hoon;Ahn, Hyung-Joon
    • Journal of Oral Medicine and Pain
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    • v.44 no.3
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    • pp.118-122
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    • 2019
  • Paraneoplastic pemphigus (PNP) is a rare and often fatal autoimmune blistering disease accompanied by both benign and malignant neoplasms. Usually, oral, skin, and mucosal lesions are the earliest manifestations shown by PNP patients. Oral ulcers are initial lesions in various autoimmune diseases like pemphigus, bullous pemphigoid, erythema multiforme, graft-versus-host, lichen planus, it does not improved despite of high-dose steroid therapy. We report a-35-year-old female who presented oral ulceration, lip crust and skin lesions. By doing several examinations, such as enzyme-linked immunosorbent assay, incisional biopsy with indirect immunofluorescence, she was diagnosed PNP with non-Hodgkin's lymphoma on pancreas.