• Title/Summary/Keyword: ELISA method

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The detection of Toxoplasma gondii ME49 infections in BALB/c mice using various techniques

  • Hae-Ji Kang;Jie Mao;Min-Ju Kim;Keon-Woong Yoon;Gi-Deok Eom;Ki-Back Chu;Eun-Kyung Moon;Fu-Shi Quan
    • Parasites, Hosts and Diseases
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    • v.61 no.4
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    • pp.418-427
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    • 2023
  • Toxoplasma gondii infections are primarily diagnosed by serological assays, whereas molecular and fluorescence-based techniques are garnering attention for their high sensitivity in detecting these infections. Nevertheless, each detection method has its limitations. The toxoplasmosis detection capabilities of most of the currently available methods have not been evaluated under identical experimental conditions. This study aimed to assess the diagnostic potential of enzyme-linked immunosorbent assay (ELISA), real-time polymerase chain reaction (RT-PCR), immunohistochemistry (IHC), and immunofluorescence (IF) in BALB/c mice experimentally infected with various doses of T. gondii ME49. The detection of toxoplasmosis from sera and brain tissues was markedly enhanced in mice subjected to high infection doses (200 and 300 cysts) compared to those subjected to lower doses (10 and 50 cysts) for all the detection methods. Additionally, increased B1 gene expression levels and cyst sizes were observed in the brain tissues of the mice. Importantly, IHC, IF, and ELISA, but not RT-PCR, successfully detected T. gondii infections at the lowest infection dose (10 cysts) in the brain. These findings may prove beneficial while designing experimental methodologies for detecting T. gondii infections in mice.

Comparison of blood parameters according to fecal detection of Mycobacterium avium subspecies paratuberculosis in subclinically infected Holstein cattle

  • Seungmin Ha ;Seogjin Kang ;Mooyoung Jung ;Sang Bum Kim ;Han Gyu Lee ;Hong-Tae Park ;Jun Ho Lee ;Ki Choon Choi ;Jinho Park ;Ui-Hyung Kim;Han Sang Yoo
    • Journal of Veterinary Science
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    • v.24 no.5
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    • pp.70.1-70.14
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    • 2023
  • Background: Mycobacterium avium subspecies paratuberculosis (MAP) causes a chronic and progressive granulomatous enteritis and economic losses in dairy cattle in subclinical stages. Subclinical infection in cattle can be detected using serum MAP antibody enzyme-linked immunosorbent assay (ELISA) and fecal polymerase chain reaction (PCR) tests. Objectives: To investigate the differences in blood parameters, according to the detection of MAP using serum antibody ELISA and fecal PCR tests. Methods: We divided 33 subclinically infected adult cattle into three groups: seronegative and fecal-positive (SNFP, n = 5), seropositive and fecal-negative (SPFN, n = 10), and seropositive and fecal-positive (SPFP, n = 18). Hematological and serum biochemical analyses were performed. Results: Although the cows were clinically healthy without any manifestations, the SNFP and SPFP groups had higher platelet counts, mean platelet volumes, plateletcrit, lactate dehydrogenase levels, lactate levels, and calcium levels but lower mean corpuscular volume concentration than the SPFN group (p < 0.017). The red blood cell count, hematocrit, monocyte count, glucose level, and calprotectin level were different according to the detection method (p < 0.05). The SNFP and SPFP groups had higher red blood cell counts, hematocrit and calprotectin levels, but lower monocyte counts and glucose levels than the SPFN group, although there were no significant differences (p > 0.017). Conclusions: The cows with fecal-positive MAP status had different blood parameters from those with fecal-negative MAP status, although they were subclinically infected. These findings provide new insights into understanding the mechanism of MAP infection in subclinically infected cattle.

Cellular Analysis and Measurement of Mucin in Sputum of Chronic Airway Disease (만성기도질환의 객담세포분석과 mucin의 측정)

  • Kim, Ki-Up;Kim, Yang-Ki;Shin, Chan-Young;Kim, Do-Jin;Uh, Soo-Taek;Kim, Yong-Hoon;Ko, Kwang-Ho;Park, Choon-Sik
    • Tuberculosis and Respiratory Diseases
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    • v.49 no.1
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    • pp.82-92
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    • 2000
  • Background : In chronic airway disease, mucus secretion is increased, but extraction of mucin, which is the main component of mucus secretion, is a very complicated and limited in clinical use. Recently, monoclonal antibody for mucin was developed for possible clinical use. In this study, cellular analysis and quantification of respiratory mucin in sputum of patients with chronic airway diseases were performed. Method : Sputum was collected from patients with asthma(n=33), bronchiectasis(n=8) or chronic bronchitis (n=13) by spontaneous expectoration or by hypertonic saline induction. Collected sputums was treated by 0.1% dithiotreitol to dissociate the disulfide bond of the mucus and filtered through a nylon gauze. Total cell count, viability and differential count were measured. For detection of mucin, collected samples were treated with sodium dodecyl sulfate polyacrylamide gel electrophoresis and then with monoclonal antibody(HMO2), as the primary antibody, and PAS stain. The amount of mucin was measured with ELISA by HMO2. Correlation with clinical information, cellular analysis, and amount of measured mucin were analyzed. Results : Total cell counts of sputum were significantly increased in patients with bronchiectasis but viability remained the same. Eosinophils were significantly increased in patients with asthma, neutrophils in bronchiectasis chronic bronchitis, respectively (p<0.05). The results of Western blotting and PAS staining confirmed the presence of glycoproteins and matched? with mucin. The amounts of mucin measured by ELISA were not significantly different among the disease groups. Significant correlation was identified between the amount of mucin and viability(r=-0.482, p<0.05). Conclusion : Inflammatory cells in the sputum of those with chronic airway disease were different for each disease type. Measurement of mucin by ELISA via monoclonal antibodies may be a simple method for the evaluation of chronic airway disease.

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Simple and Rapid Identification of Low Level Hepatitis B Virus DNA by the Nested Polymerase Chain Reaction

  • Jang, Jeong-Su;Lee, Kong-Joo
    • Archives of Pharmacal Research
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    • v.19 no.6
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    • pp.469-474
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    • 1996
  • A rapid and sensitive method has been developed to detect hepatitis B virus DNA (HBV) by nested polymerase chain reaction (PCR) technique with primers specific for the surface and core regions in capillary thermal cycler within 80 min. The lower limit for detection by present PCR method is $10^{-5}$ pg of recombinant HBV DNA which is equivalent to that determined by one round of PCR amplification and Southern blot hybridization analysis. When boiled HBV positive serum was serially diluted 10-fold, HBV DNA was successfully determined in $1{\mu}l-10^{-3}$ of serum. HBV DNA was detected by present method in 69 clinical samples including HBsAg positives and negatives by enzyme-linked immunosorbent assay (ELISA). When serum samples were amplified by nested PCR using surface and core region primers, HBV DNAs were detected in 37 of 69 samples (53.6%) and 18 of 69 samples (26.1%), respectively. These results can inform the infectious state of HBsAg positive pateints. A simple and rapid nested PCR protocol by using boiled serum as DNA template has been described for the clinical utility to determine HBV DNA in human serum.

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A simplified design approach for modelling shear force demand on tower walls supported on a transfer structure in regions of lower seismicity

  • Yacoubian, Mehair;Lam, Nelson;Lumantarna, Elisa;Wilson, John L.
    • Earthquakes and Structures
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    • v.15 no.1
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    • pp.97-111
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    • 2018
  • Buildings featuring a transfer structure can be commonly found in metropolitan cities situated in regions of lower seismicity. A transfer structure can be in the form of a rigid plate or an array of deep girders positioned at the podium level of the building to support the tower structure of the building. The anomalous increase in the shear force demand on the tower walls above the podium is a major cause for concern. Design guidance on how to quantify these adverse effects is not available. In this paper a simplified method for quantifying the increase in the shear force demand on the tower walls is presented. In view of the very limited ductile nature of this type of construction the analysis presented herein is based on linear elastic behaviour.

Development and Characterization of Anti-gliadin Polyclonal Antibody in Wheat

  • Chang, Suk Joo;Hong, Byung Hee;Seo, Yang Weon
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.44 no.4
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    • pp.339-344
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    • 1999
  • Immunological method has been applied in biochemical genetic analysis of seed storage proteins. We developed and characterized anti-gliadin polyclonal antibody (AGPab) specific to gliadin fractions whose quality and quantity were known to be associated with wheat end-use quality. Reactions of anti-gliadin polyclonal antibody (AGPab) to gliadin were linearly decreased as AGPab and antigen were diluted. Dot-blot and immunoblot assay showed that produced AGPab specifically reacted to gliadin and mainly $\alpha$-, $\beta$-, and ${\gamma}$-gliadin subunits. Enzyme-linked immuno- sorbent assay (ELISA) was applied for quantifi-cation of gliadins in Korean wheat cultivars and breeding lines by using AGPab. High reactions between AGPab and gliadins were found in wheat cultivars Olmil and Olgeurumil. Significant difference of optical densities for alcohol soluble proteins among crop species was found, as wheat showed the highest value (0.697) followed by rye (0.295), and barley (0.066).

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Semantics-aware Obfuscation for Location Privacy

  • Damiani, Maria Luisa;Silvestri, Claudio;Bertino, Elisa
    • Journal of Computing Science and Engineering
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    • v.2 no.2
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    • pp.137-160
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    • 2008
  • The increasing availability of personal location data pushed by the widespread use of location-sensing technologies raises concerns with respect to the safeguard of location privacy. To address such concerns location privacy-preserving techniques are being investigated. An important area of application for such techniques is represented by Location Based Services (LBS). Many privacy-preserving techniques designed for LBS are based on the idea of forwarding to the LBS provider obfuscated locations, namely position information at low spatial resolution, in place of actual users' positions. Obfuscation techniques are generally based on the use of geometric methods. In this paper, we argue that such methods can lead to the disclosure of sensitive location information and thus to privacy leaks. We thus propose a novel method which takes into account the semantic context in which users are located. The original contribution of the paper is the introduction of a comprehensive framework consisting of a semantic-aware obfuscation model, a novel algorithm for the generation of obfuscated spaces for which we report results from an experimental evaluation and reference architecture.

Ultra-sensitive Determination of Salinomycin in Serum Using ICP-MS with Nanoparticles

  • Cho, H.K.;Lim, H.B.
    • Bulletin of the Korean Chemical Society
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    • v.35 no.11
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    • pp.3195-3198
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    • 2014
  • An ultra-sensitive detection method for small molecules such as antibiotics was developed using ICP-MS with magnetic and $TiO_2$ nanoparticles. Since most of the antibiotics are too small to employ a sandwich-type extraction through an immunoreaction, a non-specific platform was employed, in which the target was extracted by magnetic separation, followed by tagging with $TiO_2$ nanoparticles of 11.2 nm for ICP-MS measurement. The detection limit for salinomycin obtained from spiked serum samples was $0.4ag\;mL^{-1}$ (${\pm}10.3%$), which was about $1.5{\times}10^6$ times lower than that of LC-MS/MS and about $1.2{\times}10^{11}$ times better than that of ELISA. Such an excellent sensitivity enabled us to study the toxicity of antibiotics exposed to human beings by determining them in serum.

The Effect of Cultural Conditions on the Aflatoxin Production of Aspergillus Flavus ATCC 15517 (배양조건이 Aspergillus flavus ATCC 15517의 Aflatoxin 생성에 미치는 영향)

  • 정덕화;이용욱;김용호;김성영;김종규
    • Journal of Environmental Health Sciences
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    • v.16 no.1
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    • pp.45-53
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    • 1990
  • To investigate the effect of cultural condition on the aflatoxin production of Aspergillus flayus ATCC 15517, mixed culture with Aspergillus niger, better kind of media and size of Cultural vessels were examined. YES medium was better than SLS medium for this study. Small scale test tube culture was showed the possibility to simply examine the growth, total acidity, pH and aflatoxin production during cultivation, and also could reduce the second contamination of aflatoxin B1 from large scale broth cultured. Especially ELISA method is simple, sensitive and specific and therefore well suited to small scale of test tube culture. Mixed culture significantly reduced the aflatoxin production of Aspergillus fiavus ATCC 15517 and showed almost 95% inhibition of that level during the incubtation.

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The Current Status and Future Outlook of Quantum Dot-Based Biosensors for Plant Virus Detection

  • Hong, Sungyeap;Lee, Cheolho
    • The Plant Pathology Journal
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    • v.34 no.2
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    • pp.85-92
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    • 2018
  • Enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR), widely used for the detection of plant viruses, are not easily performed, resulting in a demand for an innovative and more efficient diagnostic method. This paper summarizes the characteristics and research trends of biosensors focusing on the physicochemical properties of both interface elements and bioconjugates. In particular, the topological and photophysical properties of quantum dots (QDs) are discussed, along with QD-based biosensors and their practical applications. The QD-based Fluorescence Resonance Energy Transfer (FRET) genosensor, most widely used in the biomolecule detection fields, and QD-based nanosensor for Rev-RRE interaction assay are presented as examples. In recent years, QD-based biosensors have emerged as a new class of sensor and are expected to open opportunities in plant virus detection, but as yet there have been very few practical applications (Table 3). In this article, the details of those cases and their significance for the future of plant virus detection will be discussed.