• Title/Summary/Keyword: immuno-PCR

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Detection of Serum IgE Specific to Mite Allergens by Immuno-PCR

  • Lee, Kyung-Woo;Hur, Byung-Ung;Chua, Kaw-Yan;Kuo, I-Chun;Song, Suk-Yoon;Cha, Sang-Hoon
    • IMMUNE NETWORK
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    • v.8 no.3
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    • pp.82-89
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    • 2008
  • Background: Although a skin test is the primary option for detecting allergen-specific IgE in clinics, the serum IgE immunoassay is also important because it allows for the diagnosis of allergy without any accompanying adverse effect on the patient. However, the low detection limit of IgE levels by immunoassay may restrict the use of the method in some occasions, and improving its sensitivity would thus have a significant implication in allergy-immunology clinics. Methods: In this study, we attempted to detect specific serum IgE by using immuno-polymerase chain reaction (IPCR) which combines the antigen-antibody specificity of enzyme-linked immunosorbent assays (ELISAs) with the amplification power of PCR. Results: Our results demonstrated that Blo t5-specific serum IgE can be detected by IPCR with a 100-fold higher sensitivity than ELISA, and cross-reactivity of serum IgE to other mite allergens is able to be analyzed by using only $0.3{\mu}l$ of serum sample. Use of real-time IPCR seemed to permit more convenient determination of specific serum IgE as well. Conclusion: We believe that IPCR can serve as a valuable tool in determining specific serum IgE, especially when the amount of serum sample is limited.

Anti-allergic Effects of Shinbijen (신비전(新鼻煎)의 항알레르기 효과)

  • Kim, Kyung-Yeol;Kim, Sang-Tae;Lee, Jong-Hwa
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.21 no.6
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    • pp.1525-1533
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    • 2007
  • The discovery of drugs on the treatment of rhinitis and allergic disease is a very important subject in human heath. The Shinbijen has been used for centruries as a traditional medicine in Korea and is known to have an anti-inflammatory effect. However, its specific mechanism of action is still unknown. In this report, we investigated the effect of hot water extract from Shinbijen on OVA-mediated allergic reaction and studied its possible mechanism of action using fluorescenc microscopy and RT-PCR analysis. Shinbijen inhibited OVA-induced rhinitis, total cells in BFA and lymphocyte related to inflammation in mice. Shinbijen decreased immuno response, which activated by IL-4, COX-2 and iNOS expression. in tissue. Shinbijen reduced inflammatory molecule release from mice lung tissue and CD4/CD8 cells activated by cardiac blood. Shinbijen decreased OVA-induced IL-4 and iNOS levels in pulmonary alveoli. Our findings provide evidence that Shinbijen inhibits OVA-induced allergic reactions, and also demonstrate the involvement of inflammation and allergic disease in these effects.

Salmonella species 검출용 DNA Probe 분석시스템 고안

  • Lee, Ung-Hui;Baek, Se-Hwan
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.711-712
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    • 2000
  • DNA probe assay comprising a microwell as' solid matrix for the immobilization of streptavidin (SA) and an oligonucleotide with covalently bound fluorecein as detection probe was developed. The insolubilized SA captured the biotinylated DNA product of polymerase chain reaction (PCR), and the product was denatured under a basic condition. The remaining single-stranded DNA on the solid surface was hybridized with the probe for signal generation that was performed based on enzyme-linked immuno -reactions.

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Molecular Discrimination of Cervidae Antlers and Rangifer Antlers

  • Kim, Eun-Jin;Jung, Young-Ja;Kang, Shin-Jung;Chang, Seung-Yup;Huh, Keun;Nam, Doo-Hyun
    • BMB Reports
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    • v.34 no.2
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    • pp.114-117
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    • 2001
  • Cervi Parvum Cornu is widely used as a hemopoietic, tonifying, growth-promoting, cardiotonic, and immuno-modulating agent in Korea. In order to develop the quality control method of Cervi Parvum Cornu by the identification of the biological source or origin, the molecular approach was applied using PCR (polymerase chain reaction) and PCR-RFLF (PCR-restriction fragment length polymorphism) analysis. In the PCR analysis of the mitochondrial 12S rRNA gene and cytochrome b gene regions, no distinctive DNA bands from Cervidae (deer) antlers and Rangifer (reindeer) antlers were observed. However, when the amplified products in the mitochondrial cytochrome b gene region were subjected to restriction digestion with TaqI, Cervidae antlers showed an undigested state of 380 by band, differently from two bands of 230 by and 1S0 by from Rangifer antlers. Based on this finding, the base sequences of amplified PCR products in the range of mitochondria) cytochrome b gene from Cervidae antlers and Rangifer antlers were determined and subjected to restriction analysis by various endonucleases. The results showed that antlers from Rangifer species could be simply discriminated with other antlers from 8 Cervidae species (Chinese deer, Russian deer, Hong Kong deer, New Zealand deer, Kazakhstan deer, elk, red deer and Sika deer) by PCR-RFLP analysis using AtuI, HaeIII, HpaII or Sau3AI(MboI) as well as TaqI in the range of the mitochondrial cytochrome b gene.

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Detection of Hepatitis C Virus by RIA, Recombinant Immunoblot Assay and Nested RT-PCR (RIA, Recombinant Immunoblot Assay 및 Nested RT-PCR에 의한 C형 간염바이러스 검출)

  • Kim, Jae-Soo;Kim, Jong-Wan;Lee, Yun-Tai
    • The Journal of Korean Society of Virology
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    • v.30 no.2
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    • pp.151-159
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    • 2000
  • Hepatitis C virus (HCV) is one of the important human pathogen that can cause acute and chronic hepatitis, liver cirrhosis and hepatocellular carcinoma. Recently, the third generation radiation immuno assay (RIA) method has been developed as a very sensitive test to detect anti-HCV antibody. However, false positive is the problem with RIA test. To solve this the RIA results were compared to those of 5-antigen recombinant immunoblot assay (5-RIBA) and reverse transcription-polymerase chain reaction (RT-PCR). Among 12,767 serum samples tested from clinic visitors, total 275 (2.2%) samples were antibody positive by RIA. RIBA was performed with 148 RIA positives cases but among them was shown eighty five was antibody positive and sixty three (42.6%) was negative result. However, nested RT-PCR test was shown also carried out with 43 positive, 6 intermediates and 25 negatives of RIBA. As a result of the nested RT-PCR results, HCV antigen were detected in RIBA positive, 33.3% (2/6) RIBA intermediate and 12% (3/25). Clinical syndrome of all 148 patients as a with chronic active hepatitis (46.0%), cirrhosis (18.9%), hepatocellular carcinoma (8.1%) and others (27.0%) and they were positive in reaction by RIA test. But RIBA positive patients with 34.9% of chronic active hepatitis, 18.6% of cirrhosis, 4.6% of hepatocellular carcinoma and 41.9% of others were detected to be positive case by nested RT-PCR.

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Immuno Activation of Lectin-Conjugated Praecoxin A on IL-6, IL-12 Expression

  • Joo, Seong-Soo;Chang, Jae-Kwon;Park, Jeong-Hwan;Kang, Hee-Chul;Lee, Do-Ik
    • Archives of Pharmacal Research
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    • v.25 no.6
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    • pp.954-963
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    • 2002
  • Lectin-conjugated praecoxin A is a compound, which is combined Wheat Germ Agglutinin (WGA) Lectin with praecoxin A and also known to have an anti-tumor activity. In our lab, in order to investigate its immune reaction other than the anti-tumor activity ever known, we examined cytokines such as IL-6 and IL-12 through their mRNA expressions, which are generally secreted by macrophage both in vivo and in vitro. To analyze, we used RT-PCR for total RNAs of macrophages. As a result, we obtained that both in vitro and in vivo, lectin-conjugated praecoxin A showed an interesting increase on IL-6 and IL-12 even though it may be little hard to say the conjugated form is absolutely more effective than that of lectin or praecoxin A alone for immune response activities. Those results suggest that the conjugated form may give an additional opportunity in a future therapeutic use over its immuno activation properties.

First report of Cucumber mosaic virus in African Impatiens (Impatiens walleriana) in Korea

  • Choi, Seung Kook;Choi, Gug-Seoun;Kwon, Sun-Jung;Cho, In-Sook;Yoon, Ju-Yeon
    • Research in Plant Disease
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    • v.21 no.4
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    • pp.341-345
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    • 2015
  • Virus-like symptoms including stunt, severe mosaic with malformation of leaves, fern-like leaves and abnormal petals were observed from an African impatiens (Impatiens walleriana) grown in a plant nursery in Icheon, Korea. Serological analysis using immuno-strip kits for viruses reported in African impatiens indicated that Cucumber mosaic virus (named CMV-Im) was a causal agent for the symptomatic African impatiens. Biological properties of CMV-Im were analyzed using responses of host plant species, suggesting that CMV-Im is a typical strain that belongs to CMV subgroup I. RT-PCR analysis verified CMV-Im infection from naturally infected African impatiens or mechanically inoculated some host species. Analysis of multiple alignments of CMV capsid protein (CP) sequences showed that CMV-Im shared high CP amino acids identities with other CMV strains. Phylogenetic tree analysis for the CP sequences of CMV-Im and representative CMV strains confirmed that CMV is a typical member of CMV subgroup I. To our knowledge, it is the first report of CMV in African impatiens in Korea.

IL -12 Expression by Cefodizime As an Immuno-modulator

  • Joo, Seong-Soo;Kwon, Hee-Seung;Oh, Won-Sik;Lee, Do-Ik
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.306.1-306.1
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    • 2002
  • Cefodizime has originally been developed for treating infections as antibiotics. However. according to some of recent studies. cefodizime. a third generation cephalosporin. may potentially have the capability of stimulating chemotactic activity of neutrophils and monocytes as well as the strong immuno-modulator. In this study. we studied to learn about the expressive effect of dentritic cells and macrophage. With this background. We have studied to see if cefodizime can be a potential substance inducing an immunological function in dendritic cells and peritoneal macrophages. IL-12 activates NK cell and macrophage, and shows antiviral effect by excreting INF-${\gamma}$. In vitro. total RNAs were extracted from murine dentritic cell at 4, 8, 12, 24hr after the application of 10, 50, 100${\gamma}g$/ml of cefodizime wighout other stimulators. And we analyzed IL-12 mRNA using RT-PCR method. In conclusion. IL-12 mRNA was increased. and the results suggest that cefodizime activate TH1 cell induction, CTL differentiation as well as accelerating the increase of NK. LAK cell.

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Identification of Mycobacteria Using Polymerase Chain Reaction and Sputum Sample (객담을 이용한 Mycobacteria의 검출과 중합효소 연쇄반응의 민감성 비교)

  • Jang, Hyung Seok
    • Korean Journal of Clinical Laboratory Science
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    • v.47 no.2
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    • pp.83-89
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    • 2015
  • Although Mycobacterium tuberculosis complex strains remain responsible for the majority of diseases caused by mycobacterial infections worldwide, the increase in HIV (human immuno deficiency virus) infections has allowed for the emergence of other non-tuberculous mycobacteria as clinically significant pathogens. M. tuberculosis was detected by two-tube nested polymerase chain reaction (PCR) and non-tuberculous mycobacteria was detected by PCR-restriction fragment length polymorphism (RFLP) with Msp I. Result of niacin test is equal to result of two-tube nested PCR after culture for M. tuberculosis. In this study, acid fast bacilli stain (AFB. stain) >2+ case, Detection of Mycobacteria is similar to result before culture and after culture. AFB. stain <1+ case, result of mycobacteria is distinguished. Conclusionly, these results suggest that identification of mycobacteria must go side by side both culture and PCR for more fast and accuracy.