• Title/Summary/Keyword: Immunodiffusion

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THE CHANGES OF IMMUNOGLOBULIN ISOTYPES IN WHOLE SALIVA IN INFECTED PATIENTS OF ORAL AND MAXILLOFACIAL REGION (구강악안면 감염환자의 타액에서 Immunoglobulin Isotypes의 변화)

  • Byun, June-Ho;Chung, In-Kyo
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.26 no.2
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    • pp.186-190
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    • 2000
  • Purpose : The purpose of this study is to observe the salivary immunoglobulin level in whole saliva of infected patients and also to investigate the changes of immunoglobulin level according to its management. Materials & Methods : Thirty infected patients who have been admitted to the dept. of oral and maxillofacial surgery of Pusan National University Hospital have been selected as subjects and we analysed the changes of immunoglobulin level of $1.5{\sim}3.0ml$ of unstimulated whole saliva collected throughout four times; the day before treatment, the first day after treatment, the third day after treatment and the day before discharge. We also compared them with immunoglobulins in whole saliva that was collected from 4 normal persons as control group. In radial immunodiffusion technique with BACKMAN(Array 360 system, McLean, USA), level of immunoglobulins was analyzed. Results : The isotypes of Ig that have been found in saliva of normal persons were IgG, IgA, IgM and IgE and their mean level was 8.23, 36.41, 4.38, and 2.38 respectively. In the infected patients before the treatment, the level of IgG, IgA was remarkably higher than that of normal persons, however we could not find the difference on the level of IgM, IgE. As the infection was healing, the level of IgG, IgA was decresing significantly.

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Application of monoclonal antibody to develop diagnostic techniques for infectious bovine rhinotracheitis virus. II. Diagnosis of infectious bovine rhinotracheitis by using monoclonal antibody (소 전염성비기관염(傳染性鼻氣管炎) 바이러스에 대한 monoclonal antibody 생산(生産)과 진단법(診斷法) 개발 II. Monoclonal antibody를 이용한 소 전염성비기관염(傳染性鼻氣管炎)의 진단(診斷))

  • Jun, Moo-hyung;Kim, Duck-hwan;An, Soo-hwan;Lee, Jung-bok;Min, Won-gi
    • Korean Journal of Veterinary Research
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    • v.29 no.1
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    • pp.27-35
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    • 1989
  • To develop more specific and sensitive diagnostic methods for infectious bovine rhinotracheitis, 7-C-2 monoclonal antibody specific to polypeptides of infectious bovine rhinotracheitis virus (IBRV) was applied in indirect immunofluorescence antibody assay (IFA), indirect immunoperoxidase assay(IPA) and radial immunodiffusion enzyme assay (RIDEA). It was found that IBRV infected in MDBK cells could be detected as early as 8 hours post infection by IFA, and that IFA was more rapid and specific to identify IBRV antigen than IPA. The diagnostic efficacy of RIDEA and SN test was studied with 88 bovine sera. It was evident that RIDEA could eliminate the false positive reaction encountered in serum neutralization(SN) test, being more rapid and sensitive than the latter. Highly significant correlation coefficiency (r=0.76, p<0.01) was evaluated between the titers of sera and the diameters of RIDEA. Tracheal membranes and sera collected from 96 slaughtered cattle with lesions in respiratory organs were examined to detect IBRV antigen and antibody by IFA, RIDEA and SN test. It was presented that positive rates were 32.3% in IFA, 20.8% in RIDEA and 21.9% in SN test, and that coincidence rate between RIDEA and SN test were 100% in positive sera and 98.7% in negative sera. In conclusion, it was assumed that application of monoclonal antibody could improve the diagnostic efficacy of IBR by enhancing sensitivity and specificity of IPA, IFA and RIDEA.

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Purification of type B Staphylococcal enterotoxin (Staphylococcus aureus에서 생성된 Enterotoxin B의 분리 및 정제)

  • 이정희;신현길;김종배;한재수
    • Journal of Food Hygiene and Safety
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    • v.3 no.2
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    • pp.75-81
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    • 1988
  • Various methods such as lel-mtration on Sephadex G-SO, 75, 100 Sephacry, and Ultro gel, and lon-exchanle chromatoaraphy on Amberilte and carboxymethyl (CM)-cellulose, and Fast Protein liquid Chromatolraphy (FPLC) were applied for the purification of enterotoxin B from Staphylococcus aureus ATCC 14458 and compared one another. lon-exchanle chromatography on Amberllte resin was good enough to remove non-entrotoxln materials in culture, convlnient to use and fast although tbe purity was less tban 70%. However, CM-cellulose showed to be better purity and yield tban those of Amberilte resin. The yields of these two resins for ion-exchange cbromatograpby were about 70% and 75%, respectively. When the gel-filtration methods on Sepbadex G-50, 75, 100, Sepbacryl, and Ultro lei were applied, the purities were about 90%. FPLC was found to be tbe most efficient metbod in terms of purity (96%) and speed. For the purification of sample with large volume, particularly, tbe combined metbod, gel-mtration after Amberlite can be also used efficiently. Tbe purified toxin was found to be identical to type B enterotoxin used for reference standard by Oucbterlony immunodiffusion test.

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Standardization of an enzyme-linked immunosorbent assay for detection of antibody to avian reticuloendotheliosis virus (세망내피증 바이러스 항체검출을 위한 ELISA 표준화)

  • Sung, Haan Woo;Lee, Su Jeong
    • Korean Journal of Veterinary Research
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    • v.45 no.4
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    • pp.569-574
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    • 2005
  • Enzyme-linked immunosorbent assay (ELISA) for detection of antibodies to reticuloendotheliosis virus (REV) at single serum dilution was standardized. REV HI, one of the Korean field isolates, was inoculated into chicken embryo fibroblast (CEF) cells and was harvested from the culture fluids and cells after 10 to 12 days. Viruses were purified by centrifugation at the $107,000{\times}g$ for 12 hours on 20, 30, 45% (W/V) sucrose gradient. Virus specific fraction was collected and used as ELISA antigen. To standardize ELISA, the optimal concentration of coating antigen ($1{\mu}g/well$) and conjugate (1/1000) was determined by corrected OD (OD value of positive serum-OD value of negative serum) and P/N ratio (OD value of positive serum/OD value of negative serum). To calculate ELISA titer by measuring absorbance at 1/400 single serum dilution, serum titrations were carried out for various sample sera together with standard positive and negative sera. The observed titers of serum samples were plotted against sample/positive (s/p) ratios at 1/400 serum dilution. From the above data, the ELISA titers could be calculated by the equation of $log_{10}$ ELISA titer = 2.2763 ($log_{10}$ s/p) + 3.482 (r = 0.93). For evaluating the sensitivity, the standardized method were compared with conventional agar gel immunodiffusion (AGID) test method using serum samples collected from REV infected field chicken flocks. Fifty seven of 60 samples (95%) were positive for REV by ELISA, whereas only 11 (18.3%) samples were positive by AGID test. This results suggested that the ELISA tests developed in this study could be used for detection of antibodies to REV with high sensitivity.

Report of Four Cases of Pulmonary Pseudallescheriasis from Korea (폐(肺) Pseudallerscheria증(症) 4례(例))

  • Kim, Sang-Jae;Hong, Young-Pyo;Kim, Sin-Ok;Yoon, Young-Ja
    • The Journal of the Korean Society for Microbiology
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    • v.18 no.1
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    • pp.99-110
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    • 1983
  • Four cases of pulmonary pseudallescheriasis in patients with healed pulmonary tuberculosis are described. All four patients had a long history of antituberculous chemotherapy for pulmonary tuberculosis, but continuous sputum negativity for acid fast bacilli indicated apparent recovery from tuberculosis. They, however, complained continued intermittent hemoptysis and chronic cough. Although their chest roentgenograms did not show a clearcut mycetomal shadows in preformed cavitary lung lesions, Pseudallescheria boydii or Scedosporium apiospermum was repeatedly isolated from serial sputum specimens collected at different days for a period of over half an year or two years and their serial serum specimens produced precipitin bands with home-made antigen from 8-week old culture filtrate of P. boydii. Second fungus was isolated from sputum specimens of two patients and one was Candida albicans and the other was Aspergillus fumigatus. Sera from both patients reacted with antigens of those second fungi. Unfortunately pulmonary function of three patients did not allow surgical excision of the infected area and one patient refused surgery.

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A Survey of the Serotype Distribution of Streptococcus mutans Isolated from Dental Plaques of Caries-active and Caries-free Subjects (치아우식증환자와 치아정상인(齒牙正常人)의 치태(齒苔)에서 분리(分離)한 Streptococcus mutans의 혈청형(血淸型) 분포(分布)에 관(關)한 조사(調査) 연구(硏究))

  • Lee, Jean-Yong;Choi, Eu-Gene;Ha, Youn-Mun
    • The Journal of the Korean Society for Microbiology
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    • v.18 no.1
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    • pp.23-29
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    • 1983
  • An epidemiological survey was carried out to identify the serotypes of Streplococcus mutans isolated from carious lesions of 65 caries-active subjects(CAS) and sound tooth surfaces of 40 caries-free subjects(CFS). The autoclaved antigen extract was performed on each isolate, and then, serotypes of the isolates were determined in agar-gel immunodiffusion test. The results were as follows: 1. S. mutans was found in 78% of the samples of CAS, and of CFS. The difference of isolation frequency between CAS and CFS was not observed. 2. Only one serotype per single subject was detected in 61% of total samples, in remaining 39% of samples two or more serotypes were detected. 3. In 41.2% of CAS samples plural serotypes of S. mutans were found, whereas 35.5% of CFS samples showed plural serotypes distribution. 4. The most frequently identified serotype in each subject was serotype c; 69.5% of subjects harbored serotype c S. mutans. Serotype d was next most frequently isolated from subjects, comprising 23.2%. 5. Serotype c strain was found in 64.7% of CAS, 77.4% of CFS. 6. Of the isolates from CAS and CFS, serotype c was most commonly found, comprising 48.8%, serotype d was found in 16.3%, serotypes f. e, and g comprising 13.2%, 9.3%, and 7.8% respectively. Serotypes a and b were also found but in far lower frequencies(2.3%, 0.8%). 7. Serotype c strains were more found in CFS than in CAS, but serotypes d and e were more found in CAS.

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The Rapid Diagnosis of $\beta$-Haemolitic Streptococcus sp. by Immunoperoxidase Method (면역효소항체법에 의한 $\beta$-용혈성 연쇄구균 Streptococcus sp.의 신속진단에 대하여)

  • Chun, Seh-Kyu;Choi, Dong-Lim;Park, Soo-Il
    • Journal of fish pathology
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    • v.1 no.2
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    • pp.103-110
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    • 1988
  • For the rapid diagnosis of bacterial diseases of cultured fishes, the immunoperoxidase method was applied to the detection of $\beta$-haemolitic Streptococcus sp. strain KST-2 isolated from tilapia(Oreochromis niloticus). The suitability of field analysis and the sensitivity of the immunoperoxidase method was compared with those of the counterimmunoelectrophoresis(CIE) and the immunodiffusion(ID). Results of testing cross-reactivity which did not indicate any cross reactivity with other fish pathogens, this method was specific to Streptococcus sp. The sensitivity of this method was $1{\times}10^3CFU/ml$ which was at least $10^2$times greater than the CIE and $10^4$times greater than the ID. The immunoperoxidase method was more suitable for field application and more sensitive than other diagnostic techniques tested on this study.

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A STUDY ON THE ISOLATION AND SEROTYPING OF STREPTOCOCCUS MUTANS FROM DENTAL PLAQUE OF CARIOUS LESION (치아우식증(齒牙齲蝕症)의 환자(患者)의 우식이환부(齲蝕罹患部) 치태(齒苔)로 부터 Streptococcus mutans의 분리(分離)와 혈청형(血淸型)에 관(關)한 연구(硏究))

  • Lee, Chae-Dong;Choi, Ho-Young;Park, Sang-Jin
    • Restorative Dentistry and Endodontics
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    • v.8 no.1
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    • pp.31-44
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    • 1982
  • Streptococcus mutans strains were isolated from dental plaques of carious lesions of 53 patients on mitis-salivarius-bacitracin (MSB) and mitis-salivarius(MS) medium as a supplement. The epidemiological investigation was carried out to determine the biotypes and serotypes of S. mutans isolates. For the serotyping, autoclaved extract antigens from the isolates and serotype-specific antisera against seven known serotypes of S. mutans were prepared. The serotypes of the isolates were demonstrated in immunodiffusion test. In addtition, the prevalence of ${\beta}$-hemolysis on 5% sheep blood agar plate in restricted anaerobic condition and yellow pigment production on 5% sucrose agar plate in less anaerobic condition among the isolates were investigated. The results were as follows: 1. Forty-eight S. mutans strains were isolated from dental plaque samples of 33 patients (62.3%) among 53 patients. 2. Of the isolates, some strains were not grown on MSB medium. 3. Serotype c S. mutans was found in 60.6%, serotype d was found in 30.3% of the patients who were known to harbor S. mutans. 4. Of. the isolates, serotype c isolates were most prevalent (43.8%), serotype d isolates were 25.0%, and serotype b, e, f and g isolates were also found but in lower frequencies. Serotype a S. mutans were not detected. 5. The correlation between serotype and biotype of the isolates was found in 78.6% of the typing cases. 6. Strains revealed ${\beta}$-hemolysis were in 52.1% of the isolates, strains produced yellow pigment were in 47.9% of the isolates, and with one exception, all the strains were belong to serotype c, e and f. 7. The majority of the isolates which revealed ${\beta}$-hemolysis appeared to be yellow pigmented, these isolates were belong to serotype c, e and f.

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Immunological Comparison of Reptilian Plasma Albumins and Hemoglobins (파충류 혈장알부민 및 혈색소의 면역학적 비교)

  • Park, Sang-Yoon;Kim, Sang-Yeop;Yum, Jung-Joo
    • The Korean Journal of Zoology
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    • v.20 no.4
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    • pp.169-177
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    • 1977
  • Plasma albumin and hemoglobin were purified from the blood of Rhabdophis tigrinus. Both purified proteins and R. tigrinus crude plasma were injected into rabbits. The resulting antisera were tested for reactivity with plasma albumin and hemoglobin from, eight species of squamate and four species of non-squamate-vertebrate. Reactivity was detected qualitatively by immunodiffusion tests and immunoelectrophoreses. Antisera against plasma albumin and crude plasma reacted only with R. tigrinus plasma albumin. No antigen-antibody reactions were detected in plasma albumins of other species. Antiserum against R. tigrinus hemoglobin reacted strongly with eight squamate hemoglobins. It is likely that the reptilian plasma albumin has been experiencing rapid structural change and the reptilian hemoglobin molecules seem to be of high homogeneity. Thus, by immunological evidences, the structure of the hemoglobin molecule is considered to have been changing at slower rate than that of the plasma albumin in reptiles.

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계배 발생에 따른 닭 alpha-Fetoprotein의 분석

  • 박대규;유정아
    • The Korean Journal of Zoology
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    • v.32 no.4
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    • pp.314-321
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    • 1989
  • 배의 발생과 분화에 따른 혈청 alpha-fetoprotein (AFP)의 생리적 기능 규명을 위한 기초실험의 일환으로, 모체와는 완전히 독립된 상태에서 발생과 분화가 이루어지는 계배의 혈청을 재료로 하여 CM-Sephadex C-50, hydroxyapatite 및 DEAE-Sepharose A-25등의 크로마토그라피 방법에 의해 닭 혈청 AFP를 분리하였다. 계 AFP는 분자량이 약 60 Kd로 사람 및 다른 포유동물 AFP의 분자량 (64-74 Kd)보다 작았다. 닭 AFP, 계배혈청 및 닭 혈청 등을 토끼에 면역하여 항체를 제조하였고, 이를 이용하여 닭의 배발생 단계에 따른 혈청 AFP 함량의 변화를 전기영동 및 면역학적 방법으로 정성 및 정량 분석을 실시하였다. 혈청 AFP 함량은 발생 7일-배 (AFP 농도 : 0.81 mg/ml)부터 점차 증가하여 13일-배에서 최고치인 2.46 mgfml로 나타났으며 그 이후 급격히 감소되어 부화 직후에는 매우 낮은 농도 (0.22mg/ml)였고 부화 4일에 거의 없어 졌으며, 이와 같은 AFP 함량의 변화는 혈청 알부민 함량의 변화와 거의 반비례 함을 알았다. For the preliminary step to investigate the site of AFP synthesis, the ontogenic characteristics and the physiological function of AFP, alpha-fetoprotein was isolated from chick embryo serum through the procedures of CM-Sephadex C-50, hvdroxvapatite and DEAE-Sephadex A-50 chro-matography. The isolated AFP was proved to be pure and its molecular weight was found to be about 60 Kd. With this purified AFP, rabbit anti-chick hor was produced. The serum AFP level in chick embryo has been investagated by using polyacrylamide gel electrophoresis, immunoelec-trophoresis and single radial immunodiffusion from 7 days of incubation until 4 days after hatch-ing. It was found that the AFP level is increased from 0.81 mg/ml at daw-7 embryo to maximum value of 2.46 mg/ml at day-13 embryo, followed by rapid decreases until hatching.

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