• Title/Summary/Keyword: Immunoblot

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Immunoblot Patterns of Taenia asiatica Taeniasis

  • Jeon, Hyeong-Kyu;Eom, Kee-Seon S.
    • Parasites, Hosts and Diseases
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    • v.47 no.1
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    • pp.73-77
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    • 2009
  • Differential diagnosis of Taenia asiatica infection from other human taeniases by serology has been tested. An enzyme-linked immunoelectrotransfer blot (EITB) was applied to subjected human sera and tapeworm materials. Thirty-eight proteins reactive to serum IgG were observed between 121 and 10 kDa in adult worms, and more than 22 serum-reactive components between 97kDa and 21.5kDa were observed in eggs of T. asiatica. Antigens of adult T. asiatica revealed immunoblot bands between 120 and 21.5 kDa against T. asiatica infected sera. Antigens of adult Taenia saginata revealed 110-100, 66, 58-56, and 46 kDa immunoblot bands against T. asiatica infected sera. Antigens of adult Taenia solium also revealed 99-97, 68-66, and 46 kDa bands against T. asiatica infected sera. The immunoblot band of 21.5 kDa exhibited specificity to T. asiatica.

Estimation of HCV Test in Diagnosis for Chronic Hepatitis C Virus (만성 C형 간염바이러스 진단에 있어서 HCV검사법의 평가)

  • Jang, Soon-Mo;Yang, Byoung Seon
    • Korean Journal of Clinical Laboratory Science
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    • v.50 no.3
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    • pp.261-266
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    • 2018
  • To determine the clinical utility of an immunoblot test and RT-PCR-hybridization test, 160 samples from patients with a chronic HCV infection were analyzed by two tests. A total of 133 samples out of 150 positive samples were positive by RT-PCR-hybridization. The true positive rate of the immunoblot tests and the concordance rate of the two tests was 88.6% and 89.3%, respectively. Serotyping and genotyping were performed to evaluate the distribution of the HCV subtype in Korean isolates. HCV serotypes 1 and 2, and genotypes 1b and 2a were the most common sources of HCV infections in this group. In 49 cases studied with the serotypes and genotypes, serotypes 1 and 2 were 57.1% and 42.9%, respectively. Genotypes 1b, 1b/2b, 2a, 2a/2c, and 2b were 51.0%, 2.0%, 34.7%, 8.2%, and 4.1%, respectively. This study shows that immunoblot tests are more useful for screening HCV infections. The RT-PCR-hybridization test confirmed the HCV infection in patients with positive immunoblot test results. The serotype test is preferred over the genotype test for monitoring the progression or response to treatment. On the other hand, there were no significant differences in the response to an ${\alpha}$-interferon treatment of HCV infection with serotype type 1 or type 2 in Korea.

Protein composition and antigenicity of the tegument from Paragonimus westermani (폐흡충 총체표피의 단백질 조성 및 항원성)

  • 김석일;조승열
    • Parasites, Hosts and Diseases
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    • v.31 no.3
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    • pp.269-276
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    • 1993
  • To ascertain that tegument of Pnragonimus westermoni has specific antigenic proteins, the tegumental fraction was isolated from 10-month-old worms by 0.1% digitonin solution, and subjected to SDS-PAGE and immunoblot. Component proteins of tegumental syncytium comprised of 94, 74 (76-66), 62, 54, 44, 42, 38, 28, 26, 25, 24, 17, 15.5 and 13.5 kDa proteins. Of them, the 94, 44 and 42kDa proteins were more specific to tegument, especially the 94 kDa protein was the most prevailing one. In immunoblot, antigens of the 94, 90, 78, 76, 74, 68, 65, 63, 60, 59 and 54 kDa proteins were commonly detected by 7 sera of 10 human paragonimlasis, but none of them reacted with 5 sera of clonorchiasis. In conclusion, the 94 kDa protein was the major tegumental protein, as well as the specific antigen. The 76 and 66 kDa proteins were the minor components of tegument, which were also specific antigens of R westermcni.

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Allergenicity of Treated Chicken Egg Whites as Determined by a Passive Cutaneous Anaphylaxis Test, Immunoblot Analysis, and a Mouse Model of Food Allergy (수동피부아나필락시스 시험, immunoblot, 식품알레르기 생쥐모델에 의한 난백 처리물의 알레르기성 평가)

  • Kim, Hyun-Jung;Ryu, Ju-Hyune;Lee, Soo-Young;Shon, Dong-Hwa
    • Korean Journal of Food Science and Technology
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    • v.40 no.5
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    • pp.568-573
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    • 2008
  • The allergenicity of treated chicken egg whites (EW) was evaluated by a passive cutaneous anaphylaxis (PCA) test, immunoblot analysis, and a mouse model of food anaphylaxis. The results of the PCA test revealed that treatment with 0.3% NaOH (w/v) decreased the antigenicity of native EW to 1/4. In addition, treatment with heat ($121^{\circ}C$, 30 min) or 1% NaOH (w/v) decreased the antigenicity to 1/8 and combined treatment with 1% NaOH (w/v) and heat ($70^{\circ}C$, 15 min) decreased the antigenicity to 1/32 of that of the native EW. Immunoblot analysis revealed that the density of EW protein bands decreased in response to heat treatment, and were almost not detectable following the combined treatment. Finally, the murine model of EW anaphylaxis revealed that the mean score of systemic anaphylactic symptoms in EW challenged mice was 1.85, while the mean score in mice challenged with EW that that had been subjected to the combined treatment was only 0.20. The results of this study indicate that the most effective method of reducing EW allergenicity is combined treatment with 1% NaOH (w/v) and heat ($70^{\circ}C$, 15 min).

Identification of Allergens of Dermatophagoides farinae on Canine Atopic Dermatitis in Korea (국내 개에서 발생한 아토피성 피부염에서 Dermatophagoides farinae의 항원 확인)

  • Cho, Na-Young;Park, Seong-Jun
    • Journal of Veterinary Clinics
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    • v.31 no.6
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    • pp.490-494
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    • 2014
  • Dermatophagoides farinae plays important role in the pathogenesis of canine atopic dermatitis as environmental allergens. Also, many studies revealed that D. farinae was the main causative allergen for Korean dogs with atopic dermatitis. To identify major allergens of D. farinae in Korean atopic dogs allergic to D. farinae by immunoblot using commercial allergenic extracts, 26 dogs from two groups were enrolled in the study. Control group consists of 10 dogs with no clinical signs of disease and atopic group consists of 16 dogs diagnosed as atopic dermatitis. Sera from Korean dogs with atopic dermatitis showed six allergens of D. farinae extract by procedure of immunoblot. The molecular weights of identifying protein bands were 177, 109, 75, 44, 27, 15 kDa. The major allergens showing reactivity with greater than 50% of atopic dogs were detected at approximately 44, 109 and 177 kDa. Subsequent investigations will be carried out to verify the identity of the allergens detected in this study.

Studies on the phosphotyrsine-proteins in the rat cerbellar PSD fraction (흰쥐 소뇌 연접후치밀질내 phosphotryrosine 함유 단백질에 대한 연구)

  • 전일수;함소희;고복현
    • Journal of Life Science
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    • v.7 no.3
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    • pp.198-204
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    • 1997
  • The signal transduction through tyrosine kinases play important roles in neuronal development and synaptic regulation. We carried out immunoblot analyses to study tyrosine=phosphorylated proteins in the rat cerebellar postsynaptic density (PSD), a protein-rich cytoskeletal specialization underlying beneath the postsynaptic membrane. The overall protein composition of cerebellar PSD fractions was similar to that of the forebrain’s and only a few bands were different in Coomassie stain. Immunoblot analyses with phosphtyrosine-specific antiboy (4G10) showed that there are many more tyrosine-phosphorylated proteins in the cerebellar PSD than in the forebrain PSD. Interestiingly, a major phosphotyrosine signals in cerebellar PSD fractions was associated with a 50 kD molecular size, named as PSD-50. Migration of PSD-50 coincided with that of $\alpha$CaMKII and remained in the pellet fraction after N-octylglucoside extraction. These results indicate that tyrosine phosphorylation is important in cerebellar synaptic regulation and that the PSD-50 may be same as $\alpha$CaMKIIor a new protein which is a major substrate of tyrosine kinase.

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Changes of IgM and IgG antibody levels in experimental rabbit anisakiasis as observed by ELISA and SDS-PAGE/immunoblot (ELISA와 SDS-PAGE/immunoblot을 이용한 실험적 토끼 아니사키스증에서 혈청 항체가의 변화)

  • Yang, Hyeon-Jong;Jo, Yu-Jeong;Baek, Yeong-Han
    • Parasites, Hosts and Diseases
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    • v.29 no.4
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    • pp.389-396
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    • 1991
  • Antibody changes in experimental anisakiasis were observed in 10 rabbits which were infected each with 10 Anisakis simplex larvae. The sera were collected before and on the 6th to the 95th day after the infection. Using crude saline extract of Anisakis larvae as antigen, specific IgM and IgG antibody levels were observed by ELISA and SDS-polyacrylamide gel electrophoresislimmunoblot. Levels of specific-IgM antibody were elevated from the 6th day, reached their peaks on the lIth day after the infection, and dropped thereafter. Serum levels of IgG antibody increased from the 6th day and reached their peak on the 26th day after the infection, and decreased gradually thereafter. When SDS-PAGE of the crude extract was done, at least forty-one SDS-polypeptide bands were recognized. Of them, IgM antibody reacted mainly to the bands of 168, 95, 74, 64, 51, 47 and 34 kDa while IgG antibody reacted strongly to 168, 92, 85, 64, 58, 52, 42 and 40 kDa bands. The crude extract showed negligible cross reactions with sera of other parasitic diseases and normal control. Key words: Anisakis simplex larvae, experimental anisakiasis, rabbit, antibody, ELISA, SDS- PAGE/immunoblot.

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Immunoblot findings of calcareous corpuscles binding proteins in cyst fluid of Taenia solium metacestodes

  • Yang, Hyun-Jong
    • Parasites, Hosts and Diseases
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    • v.42 no.3
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    • pp.141-143
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    • 2004
  • After collecting calcareous corpuscles from plerocercoid of Spirometra mansoni (sparganum), we evaluated the antigenic values of calcareous corpuscles binding proteins obtained from the cyst fluid of Taenia solium metacestodes. Immunoblot analysis revealed that cysticercosis patient sera strongly recognized 10 and 95 kDa calcareous corpuscles binding proteins. This result demonstrated that calcareous corpuscles are bound with major secretory antigenic proteins, which is possibly involved in the secretory pathways of the 10 and 95 kDa proteins presenting in the cyst fluid of T. solium metacestodes.

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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Studies on the Applicability of Various Antigen Preparations in Immunoblot Diagnosis of Paragonimiasis (폐흡충증 진단에 있어서 각종 항원의 적용성)

  • Lee, Dong-Soo;Chung, Myung-Sook;Joo, Kyoung-Hwan;Rim, Han-Jong
    • Journal of agricultural medicine and community health
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    • v.17 no.1
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    • pp.46-55
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    • 1992
  • A systematic study was conducted to identify and isolate a serologically pertinent antigen with high specific activity and low cross reactivity from P.westermani antigen. Differential centrifugation of the homogenate yield three particulate and one soluble fractions ; the $480{\times}G$ pellet(Pw1), the $7650{\times}G$ pellet(Pw2) treated with n-butanol(Pw3), and $100000{\times}G$ supernatnat(Pw4). Comparison of antigenicity of these antigens, based upon differential centrifugation, to that of saline extract of P. westermani worm(SEP) was performed by SDS-PAGE and immunoblot techniques. The results obtained were as follows : 1) The ratio of absorbance value of ELISA against paragonimiasis positive pool sera to that of negative sera was highest when using Pw3 as antigen and that was lowest using Pwl. 2) Silver stained and SDS-PAGE of each antigen showed 34 and 13Kd band as common antigen band, but Pw2 didn't show clear band. 3) By immunoblot 55 and 34Kd bands using SEP and Pw4 showed strong positive reaction without cross reaction with sera from other helmenthic infections. Using Pw3, 10Kd band was observed as specific band. In conclusion, Pw3($100000{\times}G$ pellet urea soluble, treated with n-butanol) and Pw4($100000{\times}G$ supernatant) were usable for ELISA and immunoblot technique.

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