• Title/Summary/Keyword: EIA-kit

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Comparison of 5 Assays for Quantification of Antibody to Hepatitis B Virus Surface Antigen with Immunoglobulin G Preparations (면역글로불린제제 효능평가를 위한 5종 B형간염 표면항원항체검출법의 비교)

  • Shin, In-Soo;Lee, Yoo-Kyoung;Kim, Oh-Jung;Ban, Sang-Ja
    • KSBB Journal
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    • v.26 no.2
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    • pp.157-164
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    • 2011
  • Five assays for anti-HBs were compared to improve potency test of Human lgG preparations. The three commercial EIA kits were optimized including dose response curve ranges and compared by conducting a co-laboratory study. After selecting the most reproducible EIA kit, methods comparison was performed with 22 samples in 5 different days. As a result, EIA (7.7 ${\pm}$ 5.3%) and MEIA (AxSYM: 3.7 ${\pm}$ 1.9%, IMx: 1.6 ${\pm}$ 0.8%) showed precision and accuracy (100.1 ${\pm}$ 12.6%). Therefore, the validated EIA assay was established and it is believed to be comparable to current MEIA.

A study on production of early pregnancy diagnostic kit in cattle II. Production of early pregnancy diagnostic kit (소의 조기 임신진단 kit의 개발 II. 조기 임신진단 kit의 개발)

  • Kang, Chung-boo;Lee, Hyo-jong;Choe, Sang-yong
    • Korean Journal of Veterinary Research
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    • v.31 no.2
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    • pp.223-228
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    • 1991
  • Most progesterone enzyme immunoassays(EIA) are used liquid-phase double-antibody or single-antibody seperation. These methods consume considerable time and reagents because of the requirements for several washing and centrifugation steps involving the reactants. Because of these several problems, we were prompted to develop an effective enzyme-linked immunosorbent assay(ELISA) system that would be equal or superior to RIA for assay of progesterone. The results were obtained as follows. 1. Cross reaction of the progesterone antiserum with other steroids determined was shown with progesterone(100%), $11{\alpha}$-deoxycorti-costerone(2.271%), but the other steroids were shown below 0.9%. 2. Standard curve for progesterone ELISA was shown available difference according to progesterone concentration from 0 to 1,000pg/ml. 3. The lower limit of sensitivity was 0.2pg/well 4. Progesterone concentration was 1.6ng/ml for before parturition, and that was below 0.5ng/ml for after parturition. This development enzyme-linked immunosorbent assay for progesterone can be detected pregnancy diagnosis in cattle, and also applicable 10 research on physiological function including such as reproductive disorders.

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Comparison of Results According to Reaction Conditions of Thyroglobulin Test (Thyroglobulin 검사의 반응조건에 따른 결과 비교 분석)

  • Joung, Seung-Hee;Lee, Young-Ji;Moon, Hyung-Ho;Yoo, So-yoen;Kim, Nyun-Ok
    • The Korean Journal of Nuclear Medicine Technology
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    • v.21 no.1
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    • pp.39-43
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    • 2017
  • Purpose Thyroglobulin (Tg) is a biologic marker of differentiated thyroid carcinoma (DTC), produced by normal thyroid tissue or thyroid cancer tissue. Therefore, the Tg values of DTC patients is the most specific indicator for judging whether recurrence occur or whether the remaining thyroid cancer is present. Thyroid cancer is currently the most common cancer in Korea, of which 90% is differentiated thyroid cancer. The number of patients with thyroid disease of this application also increased, and an accurate and prompt results are required. However, the incubation time of the Tg commonly takes about 24 hours in our hospital, and the result reporting time is delayed, and We could not satisfied with the requirements of clinical departments and patients. In order to fulfill these requirements, experiments were conducted by shortening the incubation time between company B's Kit currently in use and company C's Kit used in other hospitals. Through these experiments, we could perform the correlation with the original method and shortening method, and could find the optimum reaction time to satisfy the needs of the departments and the patients, and we will improve the competitiveness with the EIA examination. Materials and Methods In September 2016, we tested 65 patients company B's kit and company C's kit by three incubation ways. First method $37^{\circ}C$ shaking 2hr/2hr, Second method RT shaking 3hr/2hr, Third method 1hr/1hr shaking at $37^{\circ}C$. Fourth method RT shaking 3hr method which is the original method of Company C's Kit. Fifth method, the incubation time was shortened under room temperature shaking 2hr, Sixth method $37^{\circ}C$ shaking 2hr. And we performed and compared the correlation and coefficient of each methods. Results As a result of performing shortening method on company B currently in use, when comparing the Original method of company B kit, First method $37^{\circ}C$ shaking 2hr/2hr was less than Tg 1.0 ng/mL and the ratio of $R^2=0.5906$, above 1.0 ng/mL In the value, $R^2=0.9597$. Second method RT shaking 3hr/2hr was $R^2=0.7262$ less than value of 1.0 ng/mL, $R^2=0.9566$ above than value of 1.0 ng/mL. Third method $37^{\circ}C$ shaking 1hr/1hr was $R^2=0.7728$ less than value of 1.0 ng/mL, $R^2=0.8904$ above than value of 1.0 ng/mL. Forth, Company C's The original method, RT shaking 3hr was $R^2=0.7542$ less than value of 1.0 ng/mL, and $R^2=0.9711$ above than value of 1.0 ng/mL. Fifth method RT shaking 2hr was $R^2=0.5477$ less than value of 1.0 ng/mL, $R^2=0.9231$ above than value of 1.0 ng/mL. Sixth method $37^{\circ}C$ shaking 2hr showed $R^2=0.2848$ less than value of 1.0 ng/mL, $R^2=0.9028$ above than value of 1.0 ng/mL. Conclusion Samples with both values of 1.0 ng/mL or higher in both of the six methods showed relatively high correlation, but the correlation was relatively low less than value of 1.0 ng/mL. Especially, the $37^{\circ}C$ shaking 2hr method of company C showed a sharp fluctuation from the low concentration value of 1.0 ng/mL or less. Therefore, we are planning to continuously test the time, equipment, incubation temperature and so on for the room temperature shaking 2hr method and $37^{\circ}C$ shaking 1hr/1hr of company C which showed a relatively high correlation. After that, we can search for an appropriate shortening method through additional experiments such as recovery test, dilution test, sensitivity test, and provide more accurate and prompt results to the department of medical treatment, It is competitive with EIA test.

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Anti-Wrinkle Effect of Schizandra chinensis Baillon Fermented with Lactobacillus plantarum (오미자 유산균 발효물의 주름개선 효과)

  • Lee, Jung Hee;Kim, Jong Im;Choi, Hwa Jung;Lee, Jung Hyun
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.40 no.4
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    • pp.365-371
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    • 2014
  • To identify new active anti-wrinkle ingredients, this study investigated the anti-wrinkle effects of Schizandra chinensis Baillon fermented with Lactobacillus plantarum (SCF) by assessment of cytotoxicity of human dermal fibroblast, collagen biosynthesis, matrix metalloproteinase-I (MMP-1) inhibition and elastase inhibition. S. chinensis was fermented with L. rhamnosus for 1 day at $37^{\circ}C$. The cytotoxicity of SCF was evaluated by a cytopathic effect reduction method. Effects on collagen biosynthesis and matrix metalloproteinase-I (MMP-1) of SCF were evaluated by previous reported method using procollagen type-IC peptide EIA kit and Matrix Metalloproteinase-1 Biotrack activity Assay Kit, respectively. Elastase inhibition assay was conducted by reaction of enzyme and substrate using N-Suc-$(Ala)_3$-nitroanilide as the substrate. As the results, SCF didn't show cytotoxicity against human dermal fibroblast at concentration of $100{\mu}g/mL$. Also, SCF was increased collagen synthesis and showed inhibitory effect of MMP-1 (p < 0.05). In the elastase inhibition assay, the $IC_{50}$ of SCF was $36.4{\mu}g/mL$. Therefore, our results indicated that SCF possesses anti-wrinkle effects and can be used practically for anti-wrinkle care of skin.

The Comparison of Results Among Hepatitis B Test Reagents Using National Standard Substance (국가 표준물질을 이용한 B형 간염 검사 시약 간의 결과 비교)

  • Lee, Young-Ji;Sim, Seong-Jae;Back, Song-Ran;Seo, Mee-Hye;Yoo, Seon-Hee;Cho, Shee-Man
    • The Korean Journal of Nuclear Medicine Technology
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    • v.14 no.2
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    • pp.203-207
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    • 2010
  • Purpose: Hepatitis B is infection caused by Hepatitis B virus (HBV). Currently, there are several methods, Kits and equipments for conducting Hepatitis B test. Due to ununiformed methods, it would cause some differences. To manage these differences, it needs process evaluating function of test system and reagent using particular standard substance. The aim of this study is to investigate tendency of RIA method's reagent used in Asan Medical Center through comparing several other test reagents using national standard substance. Materials and Methods: The standard substance in National Institute of Food and Drug Safety Evaluation's biology medicine consists of 5 things, 4 antigens and 1 antibody. We tested reagents using A, B company's Kits according to each test method. All tests are measured repeatedly to obtain accurate results. Results: Test result of "HBs Ag Mixed titer Performance panel" is obtained match rate compared S/CO unit standard with RIA method and EIA 3 reagents, CIA 2 reagents is that company A's reagent is 94.4% (17/18), 83.3% (15/18), B is 88.9% (16/18), 77.8% (14/18). Test result of "HBs Ag Low titer Performance panel" is obtain that EIA 2 reagents is shown 7 posive results, CIA 3 reagents is 11, and RIA method's company A's reagent is 3, B is 2 of 13 in low panel. "HBV surface antigen 86.76 IU/vial" tested dilution. A is obtain positive results to 600 times(0.14 IU/mL), B is 300 times (0.29 IU/mL). Case of "HBV human immunoglobulin 95.45 IU/vial", A is shown positive result to 10,000 times (9.5 mIU/mL) and B is 4,000 times (24 mIU/mL). Test result of "HBs Ag Working Standards 0.02~11.52 IU/mL" is shown that Company A's kit concentration level was 0.38IU/mL, company B was 2.23 IU/mL and higher level of concentration was positive results. Conclusion: When comparing various test reagents and RIA method according to National Standard substances for Hepatitis B test, we recognized that there were no significant trends between reagents. For hepatitis B virus antigen-antibody titers even in parts of the test up to 600 times the antigen, antibodies to 10,000 times the maximum positive results could be obtained. Therefore, we confirmed that results from Asan Medical Center are performed smoothly by reagents and system for hepatitis B virus test.

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Methodological validation of measuring Hanwoo hair cortisol concentration using bead beater and surgical scissors

  • Nejad, Jalil Ghassemi;Ataallahi, Mohammad;Park, Kyu-Hyun
    • Journal of Animal Science and Technology
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    • v.61 no.1
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    • pp.41-46
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    • 2019
  • Different methodologies in hair cortisol extraction may alter the final output. Thus finding the standard methodology according to a laboratory facilities is pivotal. This study was carried out to validate the feasibility of two methods of grinding hair for cortisol extraction in Korean native (Hanwoo) cattle. Hair from nine cattle including mature cows, heifers, and calves were assigned to one of the following methods for grinding hair; 1) using bead beater (BB) and 2) using surgical scissors (SS). Hair samples (> 1 g) were harvested from forehead of each individual twice (first and second measurement) to validate the results. To improve the accuracy of the obtained data, each sample was duplicated into two wells during enzyme immunoassay (EIA) analysis. Overall comparison of hair cortisol concentration (HCC) showed that the data within the range (out of the range) of standards provided by the EIA kit were 88.9% (11.1%) and 66.7% (33.3%) for BB compared with SS, respectively. In the first measurement, application of BB was tended to show higher (p = 0.056) amount of HCC compared with SS. In the second measurement application of BB showed higher (p = 0.0028) amount of HCC compared with SS. Among the cattle, calves showed higher HCC using BB compared with SS (p < 0.05). Application of BB in hair grinding methodology for Hanwoo cattle may improve cortisol extraction in comparison to application of SS method, with more consistency. Thus, it would be the preferable method to use.

Comparison of Estradiol Results according to Manufacturer (제조회사에 따른 Estradiol 결과값의 비교)

  • Kim, Dae-Woon;Shin, Hee-Jung;You, Tae-Min;Noh, Gyeong-Woon;Kim, Hyun-Joo
    • The Korean Journal of Nuclear Medicine Technology
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    • v.15 no.1
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    • pp.101-105
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    • 2011
  • Purpose: Estradiol in the menstrual cycle and ovulation induction as an important test of currently national nuclear medicine laboratory in the normal patients and patients with infertility tests are being performed. For the diagnosis of menopause is an important test with follicle stimulating hormone (FSH) and Luteinizing hormone (LH). Currently participating in external quality control of the nation's hospitals that is 54 percent of 37 hospitals, 20 hospitals have been using A's reagent. The kit's test results are highly different from other kit comes with the test results of specimens have been found. And for the phenomenon is to study the problem. Materials and Methods: Estraiol test were referred to our hospital which results of samples as more than 100pg/ml 75 specimens measured by radioimmunoassay(RIA) test with company A company B company C company D Kit, Chemiluminescent assay (CMIA) to examine and compare to the results from april to August in 2010. Results: Kit for each manufacturing company as measured by the results obtained using the average value of the correlation coefficient (R2) and A company 0.8906 B 0.9527 C 0.9547 and D company correlation coefficient of 0.873 showed a good correlation that measuring the results of A company high concentrations when Company B Company C Company D with CMIA test concentrations measured low results that the two cases were discovered specimens. Conclusion: Most of the test results of 75 samples came up with a similar trend, but two cases were reported in the patients very differently. A company result reported higher than 700 pg/ml, while the rest of other test results report was approximately 10 pg/ml. The common point of two samples more than 50 years patients are estimated to be diagnosed with cancer in postmenopausal patients receiving treatment and levels of FSH were found to be greater than 50 mIU/ml. Did not identify the exact cause. I suggest if you are using A company kit that need to again check when Estradiol result and follicle stimulating hormone results is higher.

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Ethanol Extract of Dioscorea batatas Stimulates Procollagen Production and Reduces UVB-induced MMPs Activity in Skin (마 에탄올추출물의 피부 collagen 합성 촉진 및 MMPs 활성 억제효과)

  • Kim, Dae Sung;Jeon, Byoung Kook;Lim, Nan Young;Mun, Yeun Ja;Lee, Young Eun;Woo, Won Hong
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.27 no.2
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    • pp.183-188
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    • 2013
  • Ultraviolet (UV) B irradiation induces the production of matrix metalloproteinases (MMPs), which are responsible for the degradation or synthesis inhibition of collagenous extracellular matrix in connective tissues, causing skin photoaging. In this study, we examined the inhibitory effect of MMP-1 expression of yam extract in tumor necrosis factor-alpha (TNF-${\alpha}$)-stimulated human dermal fibroblast neonatal (HDFn) cell and preventive effect of UVB-induced damage in hairless mice skin. The synthesis of procollagen and the release of MMP-1 in HDFn cells were measured by EIA kit and MMP-1 assay kit, respectively. UVB radiation was applied to the backs of the mice three times a week for 8 weeks. Mice were randomly divided into three groups, and were topical application with the Dioscorea batatas (DB, 6%) or vehicle. Reduction of TNF-${\alpha}$-induced procollagen synthesis was increased by DB (50 ug/ml), which was higher than positive control group (TGF-${\beta}$). Also, pre-treatment of HDFn cells with DB inhibited TNF-${\alpha}$-induced release of MMP-1. In vivo study, we found that preventive effect of DB against UV-induced epidermal thickness. DB suppressed the expression of MMP-3 and MMP-13 induced by UVB irradiation. Our results show that DB have preventive effect of UV-induced skin damage in hairless mice.

A Study of the Inhibitory Effect of Acer tegmentosum Max. on Fibrogenesis in Hepatic Stellate Cell Line T6 (산청목(山靑木)이 간섬유화 진행 억제에 미치는 효과에 대한 연구)

  • Lee, Seung-Bo;Woo, Hong-Jung
    • The Journal of Internal Korean Medicine
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    • v.31 no.2
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    • pp.346-355
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    • 2010
  • Objectives : This study was performed in order to investigate the anti-fibrogenic effect of Acer tegmentosum Maxim. on r at hepatic stellate cell line T6. Materials and Methods : Hepatic stellate Cells (T6) were treated with various concentrations of distilled water Acer teg mentosum Maxim. extract for 24, 48, 72 hours. After the treatment, cell viability, proliferation, procollagen levels, mRNA of AS MA, MMP-2, collagen type 1a2 and IL-6 production were measured using MTT assay, BrdU assay, RT-PCR, procollagen typ e 1 C-peptide EIA kit and murine IL-6 ELISA development kit. Results : Cell viability of HSC-T6 decreased significantly in both 24 hours and 48 hours groups in a dose-dependant man ner. Proliferation of HSC also decreased in the same way. In the RT-PCR, mRNA expression of collagen type 1a2 and ASMA decreased in the groups which were treated with Acer tegmentosum Maxim. for 24 hours. The production of procollagen tended to decrease in a dose-dependant manner in the 24 hours treated group. IL-6 production increased under Acer tegmentosum trea tment in a dose-dependant manner in both 24 and 48 hours groups. Conclusion : These results show the possibility that Acer tegmentosum Maxim. can be an effective remedy for liver fibrosi s and liver cirrhosis.

The Review of Environment, Food and Exercise on Allergy Anaphylaxis (환경, 음식 및 운동 알레르기 반응에 대한 고찰)

  • Kwak, Yi-Sub;Baek, Young-Ho;Kim, Seung-Hyun;Kim, Young-Il;Yoo, Byung-In
    • Journal of Life Science
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    • v.20 no.1
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    • pp.147-152
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    • 2010
  • Exercise-induced anaphylaxis (EIA) is a physical allergy, sometimes severe, triggered by exertion following specific food intake. It was defined for the first time in 1980. EIA is associated with different kinds of exercise. The clinical manifestations progress from itching, erythema and urticaria to some combination of cutaneous angioedema and vascular collapse. Mast cell participation in the pathogenesis of this syndrome has been proved by the findings of an elevated serum histamine level during exhaustive exercise. As predisposing factors of EIA, a specific or even nonspecific sensitivity to food has been reported. Food-dependent exercise-induced anaphylaxis (FDEIA) is a distinct form of food allergy induced by physical exercise. It is typified by the onset of anaphylaxis during exercise which was preceded by the ingestion of the causal food allergens. The diagnosis of FDEIA is heavily dependent on clinical history. Allergy tests may need to be performed using a broad panel of food and food additives. As with food allergies, FDEIA diagnosis is based on interview, biological test and skin test. Prophylaxis aims to prevent a recurrence; the patient should be given an emergency kit to deal with any recurrent episodes. After the food allergen has been identified, it should be avoided for at least 4 to 5 hours before any exercise. Two cases of EIA are presented (EIA to circumstances; FDEIA) in this paper, The diagnosis, pathophysiology and therapy of FDEIA are also reviewed.