• Title/Summary/Keyword: ELISA kit

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sICAM-1, sE-selectin, sVCAM-1 Concentration in Patients with Pulmonary Tuberculosis (폐결핵 환자에서 SICAM-1, sE-selectin sVCAM-1농도의 변화)

  • Oh, Sang-Mi;Jang, Jae-Ho;Choi, Sang-In;Lee, Heung-Bum;Lee, Yong-Chul;Rhee, Yang-Keun
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.6
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    • pp.1256-1262
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    • 1997
  • Background : Leukocyte-endothelial adhesion molecules have been implicated in the pathogenesis of inflammatory disease. ICAM-1, VCAM-1 and E-selectin are cell surface adhesion molecule on vascular endothelial cells. They are up-regulated by inflammatory cytokines and regulate the adhesion and migration of leukocytes across the endothelium. Tuberculosis, a granulomatous disorder is an infection caused by Mycobacterium tuberculosis. The clinical manifestations of tuberculosis are dependent on the cellular immune response to tubercule bacilli. Circulating adhesion molecules are probably formed by cleavage and release into the circulation of the extracellular domain of the membrane bound form. The elevated levels of circulating adhesion molecules have been reported in numerous disease state. To evaluate their role as markers of disease activity in tuberculosis, we measured a sE-selectin, sVCAM-1 and sICAM-1 levels in the serum with severities of mild, moderate and far advanced pulmonary tuberculosis. Methods : The control and test groups were divided as follows. Group I : control(n=5), Group II : patients with mild pulmonary tuberculosis(n=12), Group III : pateints with moderate pulmonary tuberculosis(n=20), Group IV : patients with far advanced pulmonary tuberculosis(n=19). Serum sICAM-1, sVCAM-1 and sE-selectin were measured by ELISA kit Results : Serum soluble adhesion molecules are elevated in patients with pulmonary tuberculosis, Circulating ICAM-1 levels were significantly elevated in patients with moderate and far advanced pulmonary tuberculosis when compared with control group. When compared with control group, serum sVCAM-1 levels showed significant elevation in patients with mild, moderate and far advanced pulmonary tuberculosis. Serum sE-selectin levels were significantly elevated in patients with far advanced pulmonary tuberculosis when compared with control group. Conclusion : These results suggest that sICAM-1, sVCAM-1, and sE-selectin may be invloved in the pathogenesis of tuberculosis. And, particularly, sICAM-1 and sVCAM-1 may be useful markers of the disease activity.

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Comparison of Two Commercial Antibody Enzyme-Linked Immunosorbent Assays for Detection of Porcine Reproductive Respiratory Syndrome Virus Infection (돼지생식기호흡기증후군(PRRS) 바이러스 감염 항체 검출 ELISA 상용 키트의 정확도 비교)

  • Pak, Son-Il;Lee, Seung-Hwan;Park, Kyung-Ae
    • Journal of Veterinary Clinics
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    • v.33 no.2
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    • pp.102-106
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    • 2016
  • More than 20 years after the first report of porcine reproductive and respiratory syndrome virus (PRRSV) in Korea, the disease is still having major impact on domestic pig health and relevant industries. Although ELISA tests are commonly used by veterinarians to guide herd management, data on diagnostic performance of the test in field settings are very limited. The objective of this study was to evaluate two commercially available PRRSV ELISA (IDEXX PRRS X3 ELISA and Bionote PRRSV ELISA 4.0) to detect antibodies against PRRSV on serum samples. To this end, a total of 1,108 sera were recruited from 35 swine farms located in Gyeonggi province and tested at the Gyeonggi Province Veterinary Service Center. All tests were performed according to the manufacturer's instructions, by laboratory technicians who routinely perform PRRS testing on blood samples. Samples were collected from two sources of swine populations with different PRRS prevalence; 60 samples (5.4%) were originated from breeding farms and the remaining 1,048 samples (94.6%) were from farrow-to-finish farms. We applied Bayesian latent class model (LCM) for two-tests in the two-population when the accuracy of the gold standard is not available. The model estimated that Bionote ELISA was a bit more specific but slightly less sensitive. The estimated sensitivity and specificity of the IDEXX ELISA were 99.8% (95% CI 98.1-100%) and 86.4% (95% CI 81.4-96.5%), respectively. Sensitivity, specificity, positive predictive value and negative predictive value for Bionote kit were 98.7% (95% CI 92.8-100%), 89.8% (95% CI 86.2-93.1%), 93.8% (95% CI 91.5-96.0%), and 97.8% (95% CI 87.1-100%), respectively. Based on the Bayesian 95% credible intervals, the sensitivity and specificity of the two ELISAs were not significantly different each other when assuming that two kits were imperfect, indicating that two kits performed equally well in terms of sensitivity and specificity in our filed setting.

Apoptosis in Human Corneal Epithelial cells induced by Exhausted Medium (Exhausted Medium에 의한 각막상피 세포의 세포고사 유도)

  • Kim, Jae-Min
    • Journal of Korean Ophthalmic Optics Society
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    • v.5 no.1
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    • pp.83-87
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    • 2000
  • To investigate exhausted-medium-induced apoptosis in human corneal epithelial(HCE) cells, this study was performed DNA gel electrophoresis, M30 CytoDEATH staining and FAS-FAS ligand ELISA. SV-40 transfected cells were grown to confluency in culture for 7days. The supernatant was harvested and filtered with $0.22{\mu}m$ filter paper. Fresh HCE cells were exposed to the filtered exhausted medium for 1~2 days. Apoptotic cells were prepared for DNA extraction and run the agarose gel for DNA ladder pattern. M30 CytoDEATH was used a tool for easy and reliable determination of very early apoptosis in HCE cells. The control and exhausted medium were assayed for soluble FAS/FAS ligand protein by ELISA. HCE cells exposed to exhausted medium showed a typical DNA ladder pattern. Sporadic M30 CytoDEATH positive cells were detected among HCE cells exposed to exhausted medium. Soluble FAS/FAS ligand levels were not elevated in the exhausted medium compared to the fresh medium control. This study suggests that possible mechanism of exhausted medium induced apoptosis does not include the FAS-FAS ligand system.

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Investigation of infection rate and genetic sequence analysis of chicken infectious anemia virus (닭 전염성빈혈 감염률 및 유전자 분석)

  • Chu, Keum-Suk;Kang, Mi-Seon;Song, Hee-Jong;Lee, Jeong-Won
    • Korean Journal of Veterinary Service
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    • v.33 no.1
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    • pp.13-21
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    • 2010
  • Chicken anemia virus (CAV) has been recognized as an immunosuppressive agent and plays role as an etiological agent of multifactorial diseases in chicken. In this study, we investigated distribution of CAV antibody by ELISA and the virus gene by PCR in poultry farms in Jeongeup, Jeonbuk province. In the test using ELISA kit, 41 (95.3%) of 43 flocks and 88.6% of the individual chickens were positive, respectively. By PCR, 90.9% of the broiler breeders and 75.0% of White-semi breeders were found positive, respectively. All hatchery was negative by PCR. Of the clinical cases from 49 poultry flocks, 87.5% of flocks and 54.7% for each samples were found positive by ELISA, respectively. By PCR test, 21 (42.9%) of 49 flocks were positive. Major clinical signs of the infected flocks were growth retardation, femoral subcutaneous bleeding, depression, limping, and continuing selection. The genetic analysis of separate N genes of CAV showed highly homologous each other. The nucleotide sequence of field isolates had homology ranged from 99.9% to 97.5% with Chinese strains, and 99.9% to 99.6% with Japanese strain. Phylogenetic analysis based on the N gene of CAV isolates showed the closely relation with Chinese strains. The results of this survey could be used as basic data for development of vaccine.

Anti-Inflammatory Effects of Rice Bran Ethanol Extract in Murine Macrophage RAW 264.7 Cells (미강에탄올추출물의 RAW264.7 세포에서 항염증효과)

  • Park, Jeong-Suk;Kim, Mi-Hye
    • YAKHAK HOEJI
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    • v.55 no.6
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    • pp.456-461
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    • 2011
  • The aim of the present study is to investigate the anti-inflammatory effect of a Rice Bran Ethanol Extract (RBE). Inflammation, such as a bacterial infection in vivo metabolites, such as external stimuli or internal stimuli to the defense mechanisms of the biological tissue a variety of intracellular regulatory factors deulin inflammatory TNF-${\alpha}$, IL-$1{\beta}$, IL-6, IL-8, such as proinflammatory cytokines, prostagrandin, lysosomal enzyme, free radicals are involved in a variety of mediators. The present study was designed to determine the effect of the RBE on pro-inflammatory factors such as NO, iNOS expression and TNF-${\alpha}$, IL-$1{\beta}$, IL-6 in lipopolysaccharide (LPS) - stimulated RAW264.7 macrophages cells. The cell toxicity was determined by MTS assay. To evaluate of anti-inflammatory effect of RBE, amount of NO was measured using the NO detection kit and the iNOS expression was measured by reverse transcriptase polymerase chain reaction (RT-PCR). And proinflammatory cytokines were measured by ELISA kit. As a result, the RBE reduced NO, iNOS expression and TNF-${\alpha}$, IL-$1{\beta}$, IL-6 production without cytotoxicity. Our results suggest that the RBE may have an anti-inflammatory property through suppressing inflammatory mediator productions and appears to be useful as an anti-inflammatory material.

Effects of Sandalwood Essential Oil on the iNOS Expression and Proinflammatory Cytokine Production (Sandalwood Essential Oil의 iNOS 발현과 염증성사이토카인의 생성에 미치는 영향)

  • Park, Jeoung-Suk;Jung, Sook-Heui
    • YAKHAK HOEJI
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    • v.57 no.1
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    • pp.70-75
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    • 2013
  • The present study was designed to determine the effect of the Sandalwood Essential Oil (Santalum album) on pro-inflammatory factors such as NO, iNOS expression and IL-$1{\beta}$, IL-6, TNF-${\alpha}$ in lipopolysaccharide (LPS) - stimulated RAW264.7 macrophages cells. The cell toxicity was determined by MTS assay. To evaluate of anti-inflammatory effect of Sandalwood Essential Oil, amount of NO was measured using the NO detection kit and the iNOS expression was measured by western blot analysis and reverse transcriptase polymerase chain reaction (RT-PCR). And proinflammatory cytokines were measured by ELISA kit. As a result, Sandalwood Essential Oil reduced NO, iNOS expression and IL-$1{\beta}$, IL-6, TNF-${\alpha}$ production without cytotoxicity. Our results suggest that the Sandalwood Essential Oil may have an anti-inflammatory property through suppressing inflammatory mediator productions and appears to be useful as an anti-inflammatory oil.

Effect of Spinal Cord Removal before or after Splitting and Washing on CNST Decontamination of Beef Carcasses

  • Lim, D.G.;Kim, D.H.;Lee, M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.11
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    • pp.1770-1776
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    • 2007
  • Beef carcasses were examined to explore the effects of spinal cord removal and washing on central nervous system tissue (CNST) decontamination of the surface during the slaughtering process. A total of 15 carcasses were split by sawing centrally down the vertebral column and left sides of split carcasses were used for analysis. Samples were collected by swabbing the surface from 4 defined parts on the interior and 4 on the exterior of carcasses from the abattoir and analyzed using an ELISA-based test. The results showed that automatic and manual spray washing decreased CNST contamination, especially on the interior ventral parts of carcass surfaces (p<0.01), but did not decrease CNST on the interior dorsal parts. Increasing washing time to 60 s did not affect the reduction of CNST contamination. Samples following spinal cord removal prior to splitting showed lower calculated levels of "risk material" than the stated limit of detection (0.1%) of the ELISA kit on interior and exterior carcass parts (p<0.01). Therefore, spinal cord removal prior to splitting could be a very effective way to minimize CNST contamination of beef carcasses.

Serosurvey for antibodies aganinst Neospora caninum in Korean indigenous cattle in the southern area of Gyeongbuk and Ulleung-gun (경북 남부지역, 울릉군 한우에서 Neospora caninum 항체 양성률 조사)

  • Lee, Jong Won;Sohn, Jun Hyung;Kim, Jung Hwa;Kim, Sang Youn;Cho, Kwang Hyun
    • Korean Journal of Veterinary Service
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    • v.41 no.3
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    • pp.185-190
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    • 2018
  • This survey was carried out to investigate the seroprevalence of Neospora caninum in Korean indigenous cattle that was raised in the southern area of Gyenongbuk and Ulleung-gun. A total of 3,162 cattle from 984 farms were tested for the presence of antibodies against N. caninum using a commercial ELISA (IDEXX Switzerland AG, Liebefeld-bern) test kit. The Seropositive ratio of cattle was 1.38% (40/2,901), 0.77% (2/261) in the southern area of Gyeongbuk and Ulleung-gun, respectively. Regional seropositive rates of farms were 5.31% (11/207), 3.45% (4/116), 1.48% (2/135), 2.13% (2/94), 2.78% (3/108), 3.91% (95/128), 4.17% (4/96), 4.35% (3/69), 6.45% (2/31) at Gumi, Yeongcheon, Gyeongsan, Gunwi, Chengdo, Goryeong, Senogju, Chilgor and Ulleung-gun, respectively. A herd of cattle under 4 years old was 1.4% (28 heads), and above the age of 5 years was 1.1% (14 heads). and in seroprevalenec cattles by herd size farms raising under 100 heads were 39 heads.

Seroprevalence of antibodies to Neospora caninum in cattle at Seoul slaughtering center (농협 서울축산물공판장 도축우에서의 Neospora caninum 혈청항체 양성률 조사)

  • Kim, Jee-Eun;Son, Jang-Won;Yang, Yun-Mo;Jeon, Hae-Chang;Jin, Kyung-Sun;Kim, Kyu-Hyun;Shin, Bang-Woo;Lee, Jung-Hark
    • Korean Journal of Veterinary Service
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    • v.34 no.2
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    • pp.179-185
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    • 2011
  • Neospora caninum is now recognized as one of the important causes of abortion in cattle worldwide, but there was no report on seroprevalence of antibodies to N. caninum in cattle at Seoul slaughtering center. Serum samples from 610 Korean indigenous cattle (Hanwoo) and 246 Holstein cattle from 452 farms located all over the country, Korea, were tested for N. caninum antibodies using a commercial enzyme-linked immunosorbent assay (ELISA) kit. Seven of 610 (1.1%) Hanwoo and 54 of 246 (22.0%) Holstein cattle were seropositive.

Effect of Theobroma cacao L. Extracts on Mouse Spleen Immune Cells Activation (카카오 열수 추출물 투여의 마우스 면역세포 활성효과)

  • Ryu, Hye-Sook
    • The Korean Journal of Food And Nutrition
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    • v.33 no.6
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    • pp.732-736
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    • 2020
  • Theobroma cacao L., a fruit of cacao trees, is a perennial plant, which belongs to Sterculiaceae, and is native to the Amazon in South Africa. It also has been known for its various biologically active effects, such as anti-oxidation, anti-cancer, and anti-bacterial. The spleen cell proliferations of mice were measured at 48 hours after treatment of Theobroma cacao L. water extracts in seven concentrations(0, 5, 10, 50, 100, 250, 500 and 1,000 ㎍/mL) an ELISA assay. The production of cytokine (IL-1β, TNF-α, IFN-γ), is secreted by macrophages stimulated with LPS, was detected by ELISA assay using the cytokine kit. From the results of in vitro study, both splenocytes and cytokine production activated by peritoneal macrophages have increased when water extracts were supplemented in the range between 250 and 500 ㎍/mL concentration. Notably, splenocytes production has a signigicant proliferation at 500 ㎍/mL concentration. The result from this research suggests that supplementation with Theobroma cacao L. water extracts may enhance the immune function by stimulating the splenocyte proliferation and improving the cytokine production activating macrophage in vitro.