• Title/Summary/Keyword: Anti-sera

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Fusobacterium nucleatum modulates serum binding to Porphyromonas gingivalis biofilm (Porphyromonas gingivalis biofilm에 대한 면역혈청의 침투력에 대한 Fusobacterium nucleatum의 조절효과)

  • Choi, Jeom-Il;Kim, Sung-Jo;Kim, Soo-Jin
    • Journal of Periodontal and Implant Science
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    • v.31 no.4
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    • pp.661-668
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    • 2001
  • Anti-P. gingivalis immune sera were obtained from mice immunized with either P. gingivalis alone, or F. nucleaturm followed by P. gingivalis. Two groups of immune sera were examined for binding capacity to P. gingivalis biofilm by confocal laser scanning microscope, Antibody avidity index was also determined for each immune sera. The results indicated that prior immunization of mice with F. nucleaturm impaired P. gingivalis-specific immune sera in binding capacity to biofilm and antibody avidity to P. gingivalis. Elevated antibody responses in patients with destructive periodontal disease has often been related to suboptimal level of protective antibody $(opsonophagocytosis)^{1-3)}$ while post-immune sera obtained with experimental animals using a single periodontal pathogen demonstrated satisfactory levels of protective function against the homologous bacterial $challenge^{4,5)}$.The reason is unclear why elevated IgG responses in periodontal patients to periodontal pathogens do not necessarily reflect their protective function. Such an immune deviation might be derived from the fact that destructive periodontal disease is cumulative result of immunopathologic processes responding to an array of different colonizing microorganisms sequentially infecting in the subgingival environmental niche. Fusobacterium nucleaturm is one of the key pathogens in gingivitis, in the transitional phase of conversion of gingivitis into destructive periodontitk, and in adult $periodontitis^{6-8)}$. It also plays a central role in coaggregation with other important microbial species in subgingival $area^{6,9,10)}$ as well as in $biofilm^{11)}$, especially with Porphyromonas gingjvalis in synergism of virulence in human periodontal disease or in animal $models^{12-14)}$. This organism has also been reported to have immune modulating activity for secondary immune response to Actinobacillus $actinomycetemcomitans^{15)}$. It is presumed that sequential colonization and intermicrobial coaggregation between intermediate and late colonizers could potentially modulate the immune responses and development of specific T cell phenotypes in periodontal lesions. We have recently demonstrated the skewed polarization of P. gingivalis-specific helper T cell clones in mice immunized with F. nucleaturm followed by P. $gingivalis.^{16)}$. Consequently F. nucleaturm may initially prime the immune cells and modify their responses to the successive organism, P. gingivalis. This could explain why one frequently observes non-protective serum antibodies to P. gingivalis in periodontal patients in contrast with those obtained from animals that were immunized with $P.gingivalis\;alone^{17)}$. The present study was performed to investigate the immune modulating effect of F. nucleatum on serum binding to experimental biofilms and the avidity of anti-P. gingivalis antibody.

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Application of Micro-BLISA in Serodiagnosis of Fascioliasis in Cattle (간질증의 효소면역학적 진단)

  • Lee, Jae-Gu;Baek, Byeong-Geol;Lee, Sang-Bok
    • Parasites, Hosts and Diseases
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    • v.23 no.1
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    • pp.95-101
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    • 1985
  • Fascioliasis in cattle is one of the most common and very serious trematode diseases in Korea. In the present study, the enzyme linked immunosorbent assay (ELISA) was applied in the diagnosis of fascioliasis using antigen of Fasciela hepatica, perokidase of conjugate anti-cattle Is G and orthophenylenediamine as a substrate by micro-method technique of Volley et at. (1976b) and MacLaren (1978) with a slight modification. Results obtained from the present study are as follows. 1. In assay for optimal dilution of stock antigen, the antigen (protein contents; 0. Bmgymz) was diluted from 1150 to 1/600 with carbonate buffer (pH 9.6), and then absorbance values were measured with 1/100 diluted sera. The regression equations between the OD values of ELISA and dilution of antigen were log Y: -0.181-0.00127X in infected sera, and log Y: -0.578-0. 000879X in normal sera. The significantly higher (p<0.05) OD value was observed in the former. 2. In assay for optimal dilution of sera, the sera were diluted from 1125 to 1/400 with in PBSJ Tween 20 (pH 7.4), and absorbance values were measured with 1/200 diluted antigen. The regression equation between the OD values of ELISA and dilution of sera were log Y: -0.1540-0.0007238X in infected sera and log Y: -0.4834-0.00116X in normal sera. The former was higher than the latter (p<0.05). 3. In the 27 cases of negative intradermal test, OD values of the ELISA are $0.447{\pm}0.144$, the 95% confidence interval (Mean+2 H SD) of the values was 0.735, and there was no case over the values. Therefore, the sensitivity of the antigen to diagnose fascioliasis was 100% in the negative case. The OD value 0.7 which is designed as a criterion (detection level of positive one) is useful for the performance of the ELISA in fascioliasis. 4. According to the OD value of criterion in the regression equations, the optimal dilutions of stock antigen and serum were 1/250 and 1/100, respectively. 5. In the 58 cases of fascioliasis from which the adult could be found in the bile ducts, the OD value was $0.846{\pm}0.224$. The 75% (44 cattle) among them had higher value with compared to the criterion, and the 60% (20 cattle) of the cases of proliferative cholangitis of 33 cattle which had been infected previousely with Fasciola sp. is higher than the criterion. 6. Prevalence of fascioliasis was 43.4% in the application of the ELISA to 272 cattle which were reared in Jeonbug district.

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Analyze of Hepatitis A Virus Genotypes in Eastern Area of Jeonnam, Korea

  • Seo, Min-Young;Lee, Hyeok-Jae;Han, Weon-Dong
    • Biomedical Science Letters
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    • v.17 no.1
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    • pp.27-37
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    • 2011
  • Hepatitis A virus (HAV) is a causative agent of triggering acute hepatitis which is transmitted by person-to-person contact and or fecal-oral route. In previous studies, most hepatitis A virus (HAV) isolates had been genotype IA in Korea. Recently, a small number of different genotypes were reported with an upsurge of acute hepatitis by HAV. Therefore, the distribution of HAV genotypes was investigated. RNA was extracted from anti-HAV IgM positive sera which were collected from February to August 2009, at a tertiary care hospital in eastern Jeonnam, Korea. Nested reverse transcription PCR and direct sequencing for VP1/P2A region of the HAV were performed. A total of 365 cases with suspected acute hepatitis were tested for anti-HAV IgM and positive results were obtained in 24 sera (9.0%), which were collected 2 to 15 days (median, 7 days) after the onset of symptoms. Of the 24 seropositive samples, 14 (58.3%) samples were positive for HAV RNA, among which 4 isolates (28.6%) were genotype IA and the other 10 (71.4%) were genotype IIIA. Both IA and IIIA genotypes were isolated from 5~6 neighboring administrative districts throughout the year without geographic or seasonal restrictions. HAV genotypes (IA and IIIA) were observed from the eastern Jeonnam for the studied.

Evaluation of Rapid IgG4 Test for Diagnosis of Gnathostomiasis

  • Wang, Yue;Ma, An;Liu, Xiao-Long;Eamsobhana, Praphathip;Gan, Xiao-Xian
    • Parasites, Hosts and Diseases
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    • v.59 no.3
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    • pp.257-263
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    • 2021
  • Human gnathostomiasis is a parasitic disease caused by Gnathostoma nematode infection. A rapid, reliable, and practical immunoassay, named dot immuno-gold filtration assay (DIGFA), was developed to supporting clinical diagnosis of gnathostomiasis. The practical tool detected anti-Gnathostoma-specific IgG4 in human serum using crude extract of third-stage larvae as antigen. The result of the test was shown by anti-human IgG4 monoclonal antibody conjugated colloidal gold. The sensitivity and specificity of the test were both 100% for detection in human sera from patients with gnathostomiasis (13/13) and from healthy negative controls (50/50), respectively. Cross-reactivity with heterogonous serum samples from patients with other helminthiases ranged from 0 (trichinosis, paragonimiasis, clonorchiasis, schistosomiasis, and cysticercosis) to 25.0% (sparganosis), with an average of 6.3% (7/112). Moreover, specific IgG4 antibodies diminished at 6 months after treatment. This study showed that DIGFA for the detection of specific IgG4 in human sera could be a promising tool for the diagnosis of gnathostomiasis and useful for evaluating therapeutic effects.

The Frequency and Distribution of Unexpected Antibodies at a Tertiary Hospital in Daejeon (대전지역 대학병원에서 동정된 비예기항체의 분포와 빈도)

  • Kang, Hee-Jung;Ihm, Chun-Hwa;Lee, Moon-Hee;Hyun, Sung-Hee;Kim, In-Sik
    • Korean Journal of Clinical Laboratory Science
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    • v.42 no.2
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    • pp.63-70
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    • 2010
  • Antibody screening and identification tests before blood transfusion are important because unexpected red antibodies can cause acute or delayed hemolytic transfusion reactions. Although a tube method was used for detecting unexpected antibodies, a column agglutination method has recently been used because of its simple procedure and a high detection of warm antibodies. This study investigated the frequency and distribution of unexpected antibodies in transfusion candidates during the recent 5 years, and transfusion characteristics in the identified cases. From January 2005 to December 2009, 46,923 sera of the cases from E hospital were screened and 98 sera were identified by the DiaMed-ID System. 272 cases (0.58%) showed positive results out of all 46,923 cases that underwent unexpected antibodies screening. Among them, unexpected antibodies were identified in 98 cases. The anti-Rh antibodies included in warm antibodies were the most frequently detected in 47 cases (47.96%). Anti-Lewis and anti-MNSs antibodies were detected in 11 cases (11.22%) and 6 cases (6.12%), respectively. Unidentified antibodies were detected in 6 cases (6.12%). Among the patients with unexpected antibodies, 43 cases (43.88%) had a history of previous transfusion. Anti-E was the most frequently detected antibody (4/14 cases, 30.77%) in the cases who had a previous history of transfusion and showed different screening results from negative to positive, This study may provide the basic data for the frequency and characteristics of red cell antibodies.

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Dot Blot Assay for Screening of Anti-hantavirus Antibodies by Using Nucleocapsid Protein of Hantaan Virus (한탄바이러스 핵단백질을 이용한 항 한타바이러스 항체 검색용 Dot Blot Assay)

  • Cho, Hae-Wol;Chung, Yeun-Jun;Kim, Chung-Lim;Ban, Sang-Ja;Nam, Jae-Hwan;Lee, Hyeong-Woo;Lee, Yoo-Jin;Kim, Eun-Jung
    • The Journal of Korean Society of Virology
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    • v.26 no.1
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    • pp.59-65
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    • 1996
  • For easy and rapid screening of hemorrhagic fever with renal syndrome (HFRS) without any laboratory equipment, dot blot enzyme immunoassay was developed and tried to detect anti-hantavirus antibodies. The nucleocapsid protein of Hantaan virus was isolated by affinity chromatography and used for making the dot strip. 28 of 29 Hantaan virus infected sera showed positive signals and 21 of 22 HFRS negative sera showed no positive signals. Anti-Seoul virus monoclonal antibody also exibited positive signal but the intensity of colorization was approximately 5 fold less than that of anti-Hantaan monoclonal antibody. The sensitivity of dot blot assay was equal or superior to indirect immunofluorescent assay (IFA) or ELISA test. Overall, the screening results with dot blot assay showed 92.2 % of concordance with IFA or ELISA test. This results suggests that dot blot assay could be applied a tool for easy and rapid screening of HFRS.

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Optimalization of ELISA using Recombinant p27 Protein of SIV for Detection of Anti-SIV (SIV의 p27 재조합 단백질을 이용한 SIV 항체 검출을 위한 ELISA의 최적 조건)

  • Kim, Eun-ok;Kim, Eun;Oh, Yoon-i;Shin, Kwang-soon;Kim, Hyun-soo;Kim, Chul-joong
    • Korean Journal of Veterinary Research
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    • v.42 no.1
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    • pp.101-108
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    • 2002
  • The p27 coding region of the SIVmac239 isolate was amplified by PCR and cloned into an expression vector, pMAL-cri, which expressed high levels of the p27 protein from Escherichia coli. The purified p27 protein was used for detection of anti-SIV antibodies with the sera from 11 macaques and 21 marmosets by immunoblot assay of which one macaque was suspicious for the SIV infection. The optimum conditions of ELISA was studied by the check board system with the recombinant purified p27 protein. For the plate coating, 200ng/well of the purified p27 was satisfactory. The conjugate was diluted 1:1000. The sera from the 32 monkeys were negative for the anti-SIV by ELISA.

Anti-hyperlipidemic Effects of Cordyceps militaris Hot-Water Extract (동충하초 열수추출액의 항고지혈증 효과에 관한 연구)

  • Kim, Han-Soo;Kim, Min-A;Jang, Seong-Ho;Lee, Won-Ki;Ryu, Jae-Young;Lee, Chun-Sik
    • Journal of Environmental Science International
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    • v.21 no.7
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    • pp.875-881
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    • 2012
  • This study was performed to observe the effects of the feeding physiological activity substance in Cordyceps militaris hot-water extract intake on the improvement of lipid components and metabolic enzyme activities in the serum of dietary hyperlipidemic rats (SD strain, male) fed the experimental diets for 5 weeks. The concentrations of cholesterol (total cholesterol, ${\beta}$-lipoprotein, LDL-cholesterol, free cholesterol and cholesteryl ester), atherosclerotic index, triglyceride (TG), phospholipid (PL) and blood glucose in sera were remarkably lower in the Cordyceps militaris extract intake group than in the hyperlipidemic rats. In the ratio of HDL-cholesterol concentration was higher percentage in the Cordyceps militaris extract intake group than in the hyperlipidemic rats. The activities aminotransferase (AST, ALT) in sera were rather lower in the Cordyceps militaris extract administration than in the hyperlipidemic rats. From the above research, Cordyceps militaris extract were effective on the improvement of the lipid compositions in sera of dietary hyperlipidemic rats.

IgG Avidity ELISA Test for Diagnosis of Acute Toxoplasmosis in Humans

  • Rahbari, Amir Hossien;Keshavarz, Hossien;Shojaee, Saeedeh;Mohebali, Mehdi;Rezaeian, Mostafa
    • Parasites, Hosts and Diseases
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    • v.50 no.2
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    • pp.99-102
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    • 2012
  • Serum samples, 100 in the total number, were collected from different laboratories in Tehran, Iran and tested for anti-Toxoplasma specific IgG and IgM antibodies using indirect immunofluorescent antibody test (IFAT). Using the IgG (chronic) and IgM (acute) positive samples, the IgG avidity test was performed by ELISA in duplicate rows of 96-well microtiter plates. One row was washed with 6 M urea and the other with PBS (pH 7.2), then the avidity index (AI) was calculated. Sixteen out of 18 (88.9%) sera with acute toxoplasmosis showed low avidity levels ($AI{\leq}50$), and 76 out of 82 (92.7%) sera in chronic phase of infection showed high avidity index (AI>60). Six sera had borderline ranges of AI. The results showed that the IgG avidity test by ELISA could distinguish the acute and chronic stages of toxoplasmosis in humans.

Detection of Human Anti-Trypanosoma cruzi Antibody with Recombinant Fragmented Ribosomal P Protein

  • Kim, Yeong Hoon;Yang, Zhaoshou;Lee, Jihoo;Ahn, Hye-Jin;Chong, Chom-Kyu;Maricondi, Wagner;Dias, Ronaldo F.;Nam, Ho-Woo
    • Parasites, Hosts and Diseases
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    • v.57 no.4
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    • pp.435-437
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    • 2019
  • Chagas disease is caused by the protozoan parasite Trypanosoma cruzi, and is endemic in many Latin American countries. Diagnosis is based on serologic testing and the WHO recommends two or more serological tests for confirmation. Acidic ribosomal P protein of T. cruzi showed strong reactivity against positive sera of patients, and we cloned the protein after fragmenting it to enhance its antigenicity and solubility. Twelve positive sera of Chagas disease patients were reacted with the fragmented ribosomal P protein using western blot. Detection rate and density for each fragment were determined. Fragments F1R1, F1R2, and F2R1 showed 100% rate of detection, and average density scoring of 2.00, 1.67, and 2.42 from a maximum of 3.0, respectively. Therefore, the F2R1 fragment of the ribosomal P protein of T. cruzi could be a promising antigen to use in the diagnosis of Chagas disease in endemic regions with high specificity and sensitivity.