• Title/Summary/Keyword: ELISA antibody

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Simultaneous Detection of Three Tobamoviruses in Cucurbits by Rapid Immunofilter Paper Assay

  • Park, Gug-Seoun;Kim, Jae-Hyun;Chung, Bong-Nam;Kim, Hyun-Ran;Park, Yong-Mun
    • The Plant Pathology Journal
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    • v.17 no.2
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    • pp.106-109
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    • 2001
  • A multi-rapid immunofilter paper assay (multi-RIPA) system was prepared for simultaneous diagnosis of three Tobamoviruses, Cucumber green mottle mosaic virus (CGMMV), Kyuri green mottle mosaic virus (KGMMV), and Zucchini green mottle mosaic virus (ZGMMV) in cucurbitaceous crops. Each of these viruses was specifically detected by the multi-RIPA from cucumber, watermelon, zucchini, and bottle gourd inoculated with the three Tobamoviruses singly or in combination. The three viruses could infect cucumber, watermelon, and bottle gourd ; however, CGMMV could not infect zucchini as the latex-coated CGMMV antibody showed a negative reaction in the multi-RIPA of the virus-infected plant extract. When the minimum detection level of multi-RIPA was compared with that of double antibody sandwich-enzyme-linked immunosorbent assay (DAS-ELISA) using CGMMV, the latter was 10 times more sensitive than the former. The detection limit of the multi-RIPA for the purified CGMMV was 50 ng/ml. In a survey of the threeviruses in cucurbits growing in commercial fields in 1999 and 2000, CGMMV was detected in watermelon and cucumber, and ZGMMV was detected only in zucchini growing in plastic houses at the suburbs of Chonju, Korea. However, KGMMV was not found in the commercially growing cucurbit crops in our study, The results suggest that the multi-RIPA can be a simple, rapid, specific and convenient tool to detect simultaneously the Tobamoviruses.

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Effect of IgY on Periodontitis and Halitosis Induced by Fusobacterium nucleatum

  • Wang, Fang;Qiao, Wu;Bao, Bin;Wang, Shujun;Regenstein, Joe Mac;Shi, Yilei;Wu, Wenhui;Ma, Ming
    • Journal of Microbiology and Biotechnology
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    • v.29 no.2
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    • pp.311-320
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    • 2019
  • Fusobacterium nucleatum is a morbific agent in periodontitis and halitosis. Egg yolk antibody (IgY) was obtained from egg yolks from chickens stimulated with F. nucleatum. This study was to assess the effectiveness of IgY on periodontitis and halitosis caused by F. nucleatum in vitro and in vivo. The growth of F. nucleatum was inhibited (p <0.05) by different concentrations of IgY in vitro and the results of a Halimeter show volatile sulfur compounds (VSCs) were reduced to $904{\pm}57ppb$ at a concentration 40 mg/ml of IgY. The changes of fatty acids of F. nucleatum were determined using GC-MS. The scores for odor index of rat saliva were decreased. The major constituent of volatile organic compounds (VOCs) including short-chain acids decreased 46.2% in 10 mg/ml IgY, ammonia decreased 70% in 40 mg/ml IgY, while aldehydes and olefine ketones were almost unchanged. The ELISA assay revealed that IL-6 and TNF-${\alpha}$ were decreased after 4 weeks' IgY treatment. Morphometric (X-ray) and histological analyses (HE) showed that IgY reduced alveolar bone loss and collagen fibers became orderly in rat models. As a result, IgY may have the potential to treat periodontitis and halitosis.

SARS-CoV-2 IgG Antibody Seroprevalence in Children from the Amritsar District of Punjab

  • Kaur, Amandeep;Singh, Narinder;Singh, Kanwardeep;Sidhu, Shailpreet Kaur;Kaur, Harleen;Jain, Poonam;Kaur, Manmeet;Jairath, Mohan
    • Korean Journal of Clinical Laboratory Science
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    • v.54 no.3
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    • pp.173-178
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    • 2022
  • The majority of the children experience milder coronavirus disease 2019 (COVID-19) symptoms. Children represent a significant source of community transmission. Children under 18 years of age account for an estimated 4.8% of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infections globally. However, no conclusive statements pertaining to the multi-fold aspects of the virus in children could be drawn due to the lower prevalence of pediatric cases. The present study was conducted to identify the indirect impact of SARS-CoV-2 infections on developing herd immunity among children in the age group 3 to 18 years by investigating their antibody levels. In the study, 240 children aged 3~18 years were recruited by the Department of Pediatrics, Government Medical College and Hospital, Amritsar, India, and quantification of the antibodies was performed at the Viral Research and Diagnostic Laboratory (VRDL), Government Medical College (GMC), Amritsar, India. Out of the 240 serum samples, 197 (82.08%) showed seropositivity, while 43 (17.92%) were seronegative. When stratified, it was observed that in the age group 3~6 years, 22.33% of children were found to have anti-SARS-CoV-2 antibodies while in the age groups 7~10 years, 11~14 years, and 15~18 years, respectively, 37.06%, 30.46%, and 10.15% were seropositive. Although there was seroconversion among children which was useful for predicting the next wave, no differences in seropositivity were observed between adults and children.

The changes of histopathology and serum anti-sparganum IgG in experimental sparganosis of mice (마우스 피하 스파르가눔증에 있어서 감염 경과에 따른 조직병리학적 병변 및 혈청 항체가의 변화 양상)

  • 홍성태;김계정
    • Parasites, Hosts and Diseases
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    • v.27 no.4
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    • pp.261-270
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    • 1989
  • The present study is intended to observe the chronologic changes of experimental sparganosis by histopathological observation and detection of circulating anti-sparganum IgG antibody using ELISA. Each of 25 mice was infected with aye spargana, and they were examined after 1, 2, 4, 10 weeks or 6 months from infection. The followings are summarized results. - 1. The plerocercoids were detected in the subcutaneous tissue of the trunk, neck or axilla, but a few often extended into the skeletal muscle. The recovery rates were 72% at the first week, 80% at the second week, 95% at the fourth week, 92% at the tenth week and 100% at .the sixth month. The larvae grew slowly in both length and weight until 6 months. 2. Histopathologically, most of the larvae were observed alive in the soft tissue or skeletal muscle. Numerous eosinophils, neutrophils, Iymphocytes and plasma cells were infiltrated focally around the worms by the second week, but they surrounded the worms to form a layer of inflammatory reaction after 4 weeks of infection. Also histiocytes and fibroblasts began to appear around the inflammatory cells at 4 weeks. After 10 weeks, the worms encircled by a thin fibrous layer were found. After 6 months, the worms were surrounded by either fibrous tissue or active inflammatory cells. The inflammation looked more severe in the tracks left by the worms, rather than around the worms. 3, The level of anti-sparganum IgG antibody in the serum showed an increase by the fourth week, and a rapid and continuous increase was observed thereafter by the tenth week after infection. The high level of the IgG antibody was maintained up to 6 months forming a plateau curve. The present results suggest that the tissue reaction and antibody production in subcutaneous sparganosis become distinctive by the fourth week after infection.

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Studios on the cell-mediated immunity in experimental Nnegleria spp. infections (실험적 Naegleria 감염에 있어서 세포매개성 면역에 관한 연구)

  • Lee, Sun-Gon;Sin, Ho-Jun;Im, Gyeong-Il
    • Parasites, Hosts and Diseases
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    • v.27 no.3
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    • pp.177-186
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    • 1989
  • Observations were made on the differences in cell-mediated immune responses in the mice infected with strongly pathogenic Naegleria fewleyi ITMAP 359, weakly pathogenic Naegzeria jadini 0400, or non.pathogenic Naegleria gruberi EGB, respectively. Variations in cell-mediated responses and changes in antibody titers according to the duration after infection wore noted. Infections were done by dropping $5{\;}{\mu}l$ saline suspension containing $10{\times}10^4$ trophozoites cultured Bxenically in the CGVS medium into the right nasal cavity of ICR mice aging about 6~7 weeks, under the anesthesia by intraperitoneal injection of'secobarbital. Following infection, delayed type hypersensitivity(DTH) iesponses in the footpad and blastogenic responses of the mouse spleen cells using [$^3H$]-thymidine were observed on the day 1, 4, 7, 10 and 14 after infection. For the preparation of amoeba Iysates, each of cultured trophosoites were homogenized with an ultrasonicator, and centrifugated at 20,000 g. The supernatants of amoeba Iysates were used as the mitogen'and antigen for ELISA. Confanavalin A(Con. A) and lipopolysaccharide(LPS) were also used as mitogens in the blastogenic response. 1. The mice infected with N, fowleri showed the mortality rate of 75.7%. The rate was 6.2% for the N. jadini infected group, while no dead mouse was observed for N. gruberi infections. 2. In regard to DTH responses in the H. fewleri infected mice, the level increased in com- parison to the control group but declined after 7 days. An increase was also noted for the JV. jadini group after 1 day, but gradual decreases were observed through the infection period. In addition, no difference was noted between the N. gruberi infected and control groups. 3. Concerning the blastogenic response of the splenocytes, it increased after 10 days in the experimental group of N, fcwleri infection, but the differences ware not statistically significant compared with control group. It was evident that N. jadini group was not different from control group either, while there was a tendency of decrease in SV. gruberi infected group. In regard to the blastogenic response of the splenocytes by LPS, it was found that the N. fowlgri, N. jadini and N. gruberi infected groups had no differences from the control group. 4. The serum antibody titer of N. fcwleri and N. jadini infected mice increased from the day 7 and 14 after infection respectively, while the N. gruberi infected mice showed no increase. In summary of the results, it was observed that there were differences in the cell-mediated immune responses and serum antibody titers in the mice infected with strongly pathogenic JV. fowleri, weakly pathogenic N. jadini, or non.pathogenic N. gruberi, respectively.

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Analysis of Immunogenicity after Hepatitis B Vaccination in Korea by Literature Review (문헌 분석에 의한 B형 간염 백신의 항체 양전률의 비교)

  • Roh, Hye Ok;Lee, Woo Gill;Sohn, Young Mo
    • Pediatric Infection and Vaccine
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    • v.5 no.2
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    • pp.245-257
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    • 1998
  • Purpose : Although hepatitis B vaccine has been available to general population in Korea since 1983, it was difficult to compare various types of hepatitis B virus(HBV) vaccines primarily due to the differences in vaccination schedule, dosage, test methods and seropositive antibody level. In this study we reviewed the results of previous studies published in Korea, which include antibody positive rates and antibody titers of various vaccines, and examined the immunogenicity of these HBV vaccines. Methods : Studies published in medical journals, university journals concerning antibody positive rates following hepatitis B vaccination were reviewed. Inclusion criteria were those studies in which seroprotective antibody rate of 10mIU/mL or the sample ratio unit of 10 RU were used as the cut-off value and in which the test methods were RIA or ELISA. Exclusion criteria were; 1) unclear or inconsistent vaccine dosage, 2) no record of antibody titers or seroconversion rate, 3) no defined antibody rate or ratio for positive rating and 4) the vaccination schedule other than 0-1-2 months or 0-1-6 months. Results : 23 out of 52 studies were subjected for the review for seroconversion rates. 1) As for the immunogenicity in each age group, the seroconversion rates of Hepaccine(Cheil Jedang) were 85.1% in infants, 83.3% in children and 62.7% in adults, indicating higher rates in infants and children compared to adults(P<0.01). The seroconversion rates of Hepavax(Korea Green Cross) were 84.7%, 81.1% and 90.8%, indicating higher rates in infants and adults compared to children(P<0.01). 2) The seroconversion rate of Hepavax was 85.6% with 0-1-6 mo. schedule, 78.5% with 0-1-2 mo. schedule with a statistically significant difference(P<0.01). 4) There was no difference of seroconversion rates between the two doses of Hepavax, $5{\mu}g$ and $10{\mu}g$ in infants and children. 5) In adults the seroconversion rates were 62.7% with Hepaccine, 90.8% with Hepavax, and 94.8% with Engerix-B(SmithKline Beecham). Conclusion : In Korea, the incidence of chronic hepatitis B is high and changing the schedule in vaccination cannot contribute to the increase of the serocoversion rate. And in order to maximize immunogenicity, more effective vaccines as well as more proper vaccination methods should be used.

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Seroepidemiological study on porcine cytomegalovirus to pigs in Korea (국내 사육 돈군내 Porcine cytomegalovirus에 관한 혈청역학적 연구)

  • Kang, Mun-il;Han, Mi;Tajima, Tomoko;Han, Dong-un;Kim, Hee-sun;Kim, Byung-han;Kim, Hong-jib;Ahn, Soo-hwan
    • Korean Journal of Veterinary Research
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    • v.38 no.4
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    • pp.756-762
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    • 1998
  • The purpose of this study was to investigate the confirmation and prevalence of porcine cytomegalovirus (PCMV) infection of pigs in Korea using enzyme-linked immunosorbent assay (ELISA). Four hundred-eighty one sera tested were collected from the areas of Kyonggi, Kangwon, Chungcheong, Cholla, Gyongsang and Cheju during the year of 1991 to 1997 except 1994. PCMV antigen, OF-1 strain, for ELISA, was prepared 19-PFT-F cell line originated from porcine fallopian tube. Positive control was used by sera made from the specific pathogen free piglets which were infected with OF-1 strain. Three hundred-sixty seven of 481 sera (76.3%) were positive against PCMV. Antibody titers of these seropositives were classified by 129 (26.8%) cases in more than 1 : 12,800, 77 (16.0%) in 1 : 6,400, 76 (15.8%) in 1 : 3,200, 44 (9.2%) in 1 : 1,600, and 41 (8.5%) in 1 : 800, respectively. Also, the seropositive pigs were divided by 87.4% (76/87) in older than 6 month-old, 73.9% (238/322) in 2~6-month old, and 73.6% (53/72) in less than 2-month old, respectively. Regional prevalence rate of PCMV infection in Korea showed 89.7% (70/78) in Chungchong, 79.8/% (83/104) in Cholla, 79.4% (143/180) in Kyonggi, 79.3% (42/53) in Gyongsang, 50% (15/30) in Kangwon, and 38.9% (14/36) in Cheju area, respectively. In the sera collected in 1991, seropositive rate was appeared as 90.2% (37/41). From 1992 to 1997 except 1994, the average infection rate to PCMV was 77.5%. Consequently, these results confirmed that PCMV in Korean piggeries was introduced at least before the year of 1991. More importantly, PCMV infection has been prevailing nation-wide in pig herds in Korea.

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Cloning and Expression of Partial Japanese Flounder (Paralichthys olivaceus) IgD

  • Choi, Dae-Han;Jang, Han-Na;Ha, Dae-Mang;Kim, Jae-Wha;Oh, Chan-Ho;Choi, Sang-Hoon
    • BMB Reports
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    • v.40 no.4
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    • pp.459-466
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    • 2007
  • The cDNA sequence of the Japanese flounder (Paralychthys olivaceus) IgD has been previously reported (GenBank accession no. AB052658) and this was followed by the detection of IgD mRNA expression in some flounder organ tissues. However, it has not been determined whether the flounder IgD gene is virtually expressed into IgD protein. To characterize the flounder immunoglobulins utilized in elucidating the mechanism, evolution and diversity of the flounder immune system, antibodies specific to IgD and IgM were necessary. In the present study, partial flounder recombinant IgD (rIgD), IgM (rIgM) and the conserved regions of IgD and IgM (rCIg) were produced by cloning the cDNA sequence using isotype specific primers which were designed to produce unique fragments of IgD and IgM specific amino acid sequences. The production of recombinant Igs was ascertained by SDS-gel electrophoresis and immunoblot analysis using anti-T7$\cdot}$Taq antibody. The produced recombinant Igs were purified using affinity columns, and used as immunogens. Antibodies specific to the isotype of flounder Igs were generated by immunizing rabbits with rfIgs and the antibodies produced were identified by enzyme-linked immunosorbent assay (ELISA) and immunoblotting. Specificities of the generated antibodies were evaluated by testing cross-reactivity between recombinant IgM and IgD. By ELISA, rabbit antibodies against the rfIgD fragment (anti-rfIgD) failed to recognize any kind of flounder serum Igs, whereas respective antibodies against rfCIg (anti-rfCIg) and rfIgM fragments (anti-rfIgM) reacted with serum Igs. Likewise, in immunoblot assays, though anti-rfIgD did not, both anti-rfCIg and anti-rfIgM bound with the ~85 kd flounder IgM heavy chain. By flow cytometry analysis, anti-rfCIg, anti-rfIgD and anti-rfIgM reacted with 6%, 3% and 6.5% of cells, respectively, suggesting that flounder IgD is not secreted in serum but expressed on flounder B-like cell surfaces as in mammals. Antibodies produced against recombinant flounder Igs could be used to develop sandwich assay systems for detecting flounder Igs and for further investigating the flounder immune system.

Prevalence of Antibodies to Human Herpesvirus 8 in Children (소아의 항 Human Herpesvirus 8 항체 양성률)

  • Han, Tae Hee;Chung, Ju Young;Kim, Sang Woo
    • Pediatric Infection and Vaccine
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    • v.12 no.2
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    • pp.108-113
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    • 2005
  • Purpose : Human herpsevirus 8(HHV-8), a gamma herpsevirus, was initially identified from Kaposi sarcoma(KS) lesions and has been known to be associated with several malignancies including Kaposi sarcoma. HHV-8 seroprevalence is variable by different geographic areas and populations. The prevalence of HHV 8 infection in Korean children is unclear. So, we investigated the prevalence of HHV-8 specific antibodies in healthy children in Seoul, Korea. Methods : Sera were obtained from 112 children(age 1~15 years, 64 males and 48 females) who visited our hospital for routine health checkup and used for investigating sero-prevalence of anti-HHV-8 antibodies. An indirect immunofluorescent assay was used to detect the IgG antibodies to the lytic viral antigen(Biotrin, Dublin, Ireland). A peptide mix ELISA kit was used to detect the IgG antibodies to peptides specific for HHV-8 open reading frame (ORF)(Biotrin, Dublin, Ireland). Results : Of 112 children, 4 children younger than 6 years of age were seropositive to HHV-8[all 4(3.5%) were positive by IFA and 2(1.8%) were positive by ELISA]. Conclusion : These results suggest that the prevalence of antibody to HHV 8 in children in Korea is very low.

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

  • Cho, Young-Shik;Yu, Seung-Shin;Ha, Gun-Woo;Lee, Sang-Gook;Cho, Myung-Hwan;Shin, Hyung-Sik;Kim, Sun-Young
    • The Journal of Korean Society of Virology
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    • v.28 no.1
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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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