• 제목/요약/키워드: ELISA system

검색결과 327건 처리시간 0.022초

Levels of common salivary protein 1 in healthy subjects and periodontal patients

  • Heo, Seok-Mo;Lee, Sol;Wang, HongTao;Jeong, Jeong Hyeok;Oh, Sang Wook
    • Journal of Periodontal and Implant Science
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    • 제46권5호
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    • pp.320-328
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    • 2016
  • Purpose: Human saliva, as a vital part of the immune defense system, contains a number of distinct proteins and peptides. Recently human common salivary protein 1 (CSP1) has been identified as an abundant salivary protein and may play a role in promoting the binding of cariogenic bacteria to salivary pellicles. However, nothing else is known regarding the role of CSP1 in periodontology. The aim of this study was to quantify and compare CSP1 levels between healthy subjects and periodontal patients. Methods: This controlled clinical study was conducted in periodontally healthy individuals and patients with chronic periodontitis Chonbuk National University Hospital, with Institutional Review Board approval. Whole saliva samples were collected from 36 healthy subjects and 33 chronic periodontitis patients and analyzed. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immune blotting were conducted to ensure that anti-CSP1 monoclonal antibody (mAb) binds to CSP1 in human saliva. A sandwich enzyme-linked immunosorbent assay (ELISA) system was house-fabricated using mAb-hCSP1#14 and mAb-hCSP1#4 as a capture and a detector mAb, respectively. The CSP1 concentrations in saliva from 36 healthy subjects and 33 periodontal patients were quantified using the CSP1 sandwich ELISA system, and the results were analyzed using the Student's t-test. Results: Immunoblot analysis using mAb-hCSP1 as a probe confirmed that CSP1 in human saliva existed as a single band with a molecular weight of approximately 27-kDa. The quantification of CSP1 concentrations by CSP1 ELISA showed that the median values (25th to 75th percentiles) of periodontal patients and healthy subjects were 9,474 ng/mL (range, 8,434.10,139 ng/mL) and 8,598 ng/mL (range, 7,421.9,877 ng/mL), respectively. The Student's t-test indicated the presence of a statistically significant difference between the 2 groups (P=0.024). Conclusions: The presence of a significant difference in CSP1 levels between healthy subjects and periodontal patients suggests that CSP1 may be a potential biomarker for the detection or screening of periodontitis patients.

HIV-1 O형 항체 진단시료의 개발 (Development of Test System for Detection of Antibody to Human Immunodeficiency Virus Type 1 Subtype O)

  • 조영식;유승신;하건우;이상국;조명환;신형식;김선영
    • 대한바이러스학회지
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    • 제28권1호
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    • pp.31-38
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    • 1998
  • In Korea, all domestic made test systems for detecting antibodies in HIV-1 contain the antigens from human immunodeficiency type 1 (HIV-1) subtype B. However, because HIV-1 subtype O is significantly different in amino acid sequences from all other subtypes of HIV-1, there has been a need for developing a test for detecting antibodies in subtype O. For this purpose, the entire nucleotide sequence corresponding to the extracellular domain of the transmembrane glycoprotein of HIV-1 subtype O was synthesized with consideration of Escherichia coli condon usage. Various regions of the extracellular domain were cloned into E. coli expression vectors and tested for levels of protein production. The nucleotide sequence, named ECTM, that can encode a 129 amino acid-long peptide, was found to be expressed at a high level in E. coli. The protein of approximately 17 kDa specifically reacted with sera from individuals infected with HIV-1 subtype O. The ECTM protein was purified to near homogeneity by the CM-T gel chromatography, using concentrated, denatured inclusion bodies. In Western blot analysis, the purified viral antigen reacted with sera from individuals infected with subtype O more efficiently than subtype B. The enzyme linked immunoabsorbent assay (ELISA) system was developed using the subtype O viral protein and compared with the commercially available kit lacking the antigens from subtype O. The ELISA kit containing the subtype O antigen ECTM alone efficiently reacted with sera from individuals infected with subtype O. The subtype O antigen-containing kit produced a positive absorbence even when sera were diluted 512-fold, suggesting a high sensitivity. The commercially available kit also reacted with subtype O sera, but produced a negative result at a dilution of 8-fold. Our results suggest that the currently available kit may not be able to efficiently detect subtype O sera and that the viral protein developed in this study may be added to the current system to maximize the detection of sera from individuals infected with subtype O.

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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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    • 제20권11호
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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.

돼지 옴 감염증 진단을 위한 바이오센서 연구 (A Biosensor for Diagnosis of Swine Sarcoptic Mange)

  • 조한근;지차호
    • Journal of Biosystems Engineering
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    • 제30권5호
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    • pp.306-311
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    • 2005
  • In this study, a biosensor was developed to rapidly diagnose the swine sarcoptic mange (Sarcoptes scabies var. suis). The ELISA was modified to reduce the processing time for rapid diagnosis. The biosensor consists of a biological reaction part, and a measurement and control part. The biological reaction part was designed for using micro-pumps and valves for fluid transportation, and the measurement control part composed of a photodiode, a light-emitting diode fur light measurement, and a microcomputer to implement assay A polystyrene covet was used as a reaction chamber. Signal output was read as the rate of change in optical density at 645nm. Eighteen pigs diagnosed with sacroptic mange and 19 control pigs were tested. Fifteen sacbies-infested pigs showed positive results ($83.3\%$ sensitivity). Sixteen control pigs showed negative results ($84.2\%$ specificity). The system could execute a diagnosis cycle in about 45 min. The results suggest that this biosensor is useful for the rapid diagnosis of swine sacroptic mange.

Immunoglobulin productivity assay를 이용(利用)한 홍삼투여(紅蔘投與) 실험동물(實驗動物)의 IgG, IgM, IgA 비교(比較) 연구(硏究) (The Comparative Study of IgG, IgM, and IgA in Laboratory Animal Administrated Red-ginseng, Using Immunoglobulin Productivity Assay)

  • 이범준;소형진;김재완;류재환
    • 대한한방내과학회지
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    • 제28권4호
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    • pp.886-895
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    • 2007
  • Objective : The immune system is a complex of systems, all of which work together to clear infection from the body. In Korea, red ginsenghas been one of the herbs most widely used to enhance the immune system for thousand of years. More recently, red ginseng has been reported to have many positive effects on the immune system. The purpose of this study was evaluate the effects of Korean red ginseng and Chinese red ginseng on IgG, IgM, and IgA, using immunoglobulin productivity assay. Methods : Male SD rats were separated into 3 groups. We administered Korean red ginseng (KRG) to one group and Chinese red ginseng (CRG) to another, with normal saline for the Control group consecutively and orally for 3 months. The dose of red ginseng was 500mg per day, as a powder with soluble water. Immunoglobulin levels from spleen cell were estimated by ELISA kit. Results : In immunoglobulin productivity assay (cell), the IgG level of the KRG group significantly increased but there was no significant difference in the IgG of the CRG group. The IgM level of the KRG group significantly increased stimulated with PWM. When it was unstimulated, the level of IgM in KRG and CRG increased together. The IgA level of the KRG group significantly increased when it was stimulated with PWM and unstimulated. Conclusion : According to the above results, oral administration of red ginseng for 3 months is considered useful for immunomodulatory effect, and Korean red ginseng may be superior to Chinese red ginseng in that effect.

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인삼(人蔘)이 중추신경계 손상 동물 모델의 재생에 미치는 영향 (The Effect of Ginseng Radix on Regeneration After Central Nervous System Injury)

  • 문형철;김윤욱;송봉근
    • Journal of Acupuncture Research
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    • 제24권6호
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    • pp.137-148
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    • 2007
  • Objectives : Following central nervous system (CNS) injury, inhibitory influences at the site of axonal damage occur. Glial cells become reactive and form a glial scar, know as gliosis. As well,myelin debris such as MAG inhibits axonal regeneration. Astrocyte-rich gliosis relates to up-regulation of GFAP and CD81, and eventually becomes a physical and mechanical barrier to axonal regeneration. It is postulated that when the astrocytic reaction is absent, regeneration of axons can occur. It was reported that treatment with anti CD81 antibodies enhanced functional recovery in rats with spinal cord injury. Methods : MAG is one of several endogenous axon regeneration inhibitors that limit recovery from central nervous system injury and disease. It was reported that molecules which block such inhibitors enhanced axon regeneration and functional recovery. Results : In this current study, the author investigated the effect of the water extract of Ginseng Radix on the regulation of CD81, GFAP and MAG which increases when gliosis occurs. MTT analysis was performed to examine cell viability, and cell based ELISA, Western Blot and PCR were used to detect the expression of CD81, GFAP and MAG. Immunohistochemistry was also performed to confirm in vivo. Conclusions : We observed that Ginseng Radix significantly down-regulates the expression of CD81, GFAP and MAG by means of cell based ELISA, Western Blot and PCR. In immunohistochemistry, expression of CD81, GFAP and MAG also decreased. Taken together, these results suggest that Ginseng Radix can be a candidate for regenerating CNS injury.

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Saccharomyces cerevisae에서 한국산 겨우살이 유래 lectin A 및 B 유전자의 발현 (Expression of Recombinant Korean Mistletoe(KM) Lectin and B genes in Saccharomyces cerevisiae)

  • 최윤혁;김종배;양웅석;황철원
    • 생명과학회지
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    • 제14권5호
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    • pp.840-846
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    • 2004
  • 본 연구는 한국산 겨우살이 lectin유전자 (A 및 B chain) 을 효모 (Saccharmyces cerevisiae) 에 형질 전환시키는 시스템을 사용한 것으로, 효모내 효과적인 lectin유전자 발현을 위하여 유전자 상에 Kozak translation initiation sequence를 PCR을 이용 삽입, 변형시켜 재 클로닝 하였다. 변형된 lectin A 및 B 유전자를 포함하는 재조합 플라스미드는 S. cerevisiae INVSc (MATa, his3 $\Delta$l, leu2, trpl-289, ura3-52) 에 형질전환 되었다. 형질전환된 효모는 ABI 3700 system을 이용한 DNA 염기서열 분석을 통해 확인되었고 재조합 한국산겨우살이 lectin 발현을 위해 2% galactose를 사용하여 유도발현되었다. 재조합 lectin A 및 B 단백질은 SDS-PACE 및 western blotting 분석을 수행한 결과 약 29kDa 크기로 확인되었다. 재조합 lectin은 세포내 가용성 단백질 1mg중 1.24∼l.75 $\mu\textrm{g}$ 수준으로 발현되어짐을 ELISA 분석을 통해 확인하였다. 한편 lectin 유전자는 galartose 유도발현 후 36시간이 되 었을 때 발현량이 최대가 되었으며 lectin A 유전자의 경우 48 시간 이후에는 발현이 억제되었다.

생쥐의 면역계에 미치는 인삼 사포닌의 영향 (The Effect of Ginseng Saponin on the Mouse Immune System)

  • 김미정;정노팔
    • Journal of Ginseng Research
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    • 제11권2호
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    • pp.130-135
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    • 1987
  • 인삼 saponin이 면역작용에 미치는 영향을 알아보기 위하여 생쥐에 단백질 항원(암닭의 ${\gamma}$-globulin)으로 면역시킨 후 1차 면역후 10일, 2차 면역후 10일에 각각 채혈하여 혈청내의 항체가를 ELISA(enzyme linked immunosorbent assay) method로 측정하였고, 또한 같은 항원으로 면역시킨 생쥐에 면역억제제를 사용하여 생쥐의 면역제를 억제시킨 후 그 회복에 미치는 영향을 알아보기 위하여 같은 방법으로 항체가를 측정하였다. 인삼 saponin을 투여한 실험군(10mg/ kg/day)은 개체에 따라 약간의 차이는 있었으나 같은 조건의 생리식염수 투여군보다 각각 훨씬 높은 항체가를 나타내었고, 면역억제제에 의한 면역억제의 회복에 있어서도 유의성있는 회복효과를 나타내었다. 따라서 면역작용에 미치는 인삼saponin의 영향은 정확한 기작은 밝혀지지 않았으나 인삼 saponin이 혈청단백질 합성을 증가시키는 효과와 함께 일종의 면역자극제(immunostimulator)로 작용하고 있는 것으로 생각된다.

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Binding Affinities of Carbohydrate-Conjugated Chlorins for Galectin-3

  • Park, Young K.;Bold, Bayarmaa;Cui, Bing Cun;Bai, Jin Quan;Lee, Woo-Kyoung;Shim, Young-Key
    • Bulletin of the Korean Chemical Society
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    • 제29권1호
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    • pp.130-134
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    • 2008
  • Carbohydrate-conjugated chlorins were synthesized for use as biosensors for the detection of the galectin-3 cancer marker. We used ELISA, SDS-gel electrophoresis, and Bradford assays to examine the binding of galectins to d-(+)-galactose- and b-lactose-conjugated chlorins. The binding affinities of these conjugated chlorins for galectin-3 were quantified using fluorescence spectroscopy. The fluorescence emission of the carbohydrate-conjugated chlorins decreased as the amount of galectin-3 in the binding reaction increased over a limited concentration range, indicating that carbohydrate-conjugated chlorins are potentially useful fluorescence biosensors for the galectin-3 cancer marker.

살충제 Imidacloprid 잔류물의 신속한 측정을 위한 생물반응 및 계측제어 시스템 개발 (Development of a Biological Reaction and Measurement Control System for Rapid Detection of the Insecticide Imidacloprid Residues)

  • 임종근;조한근
    • Journal of Biosystems Engineering
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    • 제30권2호
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    • pp.114-120
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    • 2005
  • In this study, a biological reaction and measurement control system was developed to rapidly measure the insecticide imidacloprid residues in agricultural products. The biological reaction part of the system was designed to include micro-pumps and valves for fluid transport, and a polystyrene covet as a reaction chamber. The measurement control part of the system consisted of a photodiode with a light-emitting diode for optical density measurement, and a control microcomputer to implement assay. Signal output was read as the rate of change in optical density at 645 nm. The sensitivity of the system was 2.2 ng/mL ($IC_50$). The system could execute a measurement cycle in about 19 minutes. Research will be continued to develop an automatic sampler fur imidacloprid residues from agricultural products.