• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제26권2호
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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.

• 대한수의학회지
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• 제29권1호
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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.

• 한국식품위생안전성학회지
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• 제3권2호
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• pp.75-81
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• 1988

본 실험은 Staphylococcus aureus 로부터 생성되는 enterotoxi B 의 분리 및 정제를 위하여 각종 분리방법을 비교 조사하였다. 배지로부터 Entrotoxin B를 추출하는 방법중 Amberlite CG-50 수지가 가장 간편하고 빠른 방법이었고 CM 수지는 Amberlite 수지에 비해 용출력이 떨어졌으며 분리할 수 있는 toxin의 양은 적었으나 정제도에 있어서는 약 75%로 toxin을 분리하는 처음 단계로서는 높은 편이었다. CM column을 Gradient 용출법으로 사용했을 때에는 하나의 column을 사용해 분리한 분리물 중 정제도가 85%로 가장 높았고, 용출 buffer의 농도 폭을 넓히는 것이 정제도를 높이기 위한 바람직한 방법이었다. 이 실험에 사용한 Sephadex G-50 , 75, 100, Sephacryl, Ultro gel 등의 gel filtration 방법 중 Ultro gel 에 의한 분리방법이 정제도에 있어서는 가장 우수했으며, 이온 교환 수지를 먼저 사용한 분리물에서는 모두 90% 이상의 toxin을 얻을수 있었지만, 한번의 분리도 거치지 않은 배지는 분리도와 정제도에서 현저히 떨어졌고, Sephadex G-50 은 gel colunm중 정제도가 가장 낮았다. FPLC는 위의 분리 ·정제 방법중 가장 빠른 방법이며, 적은 양의 시료로도 측정이 가증하였고, 정제도는 95% 이상이었다.

• 대한수의학회지
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• 제45권4호
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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.

• 대한미생물학회지
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• 제18권1호
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• pp.99-110
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• 1983

저자 등은 결핵으로 인해 생긴 공동을 가진 4명의 남자 환자에 있어서 성공적인 결핵화학요법으로 결핵은 치유되어 객담검사에서 결핵균은 더이상 검출되지 않았으나, 계속되는 각혈로 진균검사를 실시한 결과 Pseudallerscheria boydii(불완전세대 Scedosporium apiospermum)에 의한 진균종(眞菌腫)임이 밝혀졌다. 여러 차례의 객담검사로부터 6개월 또는 2년 이상 계속 동일한 균종이 불리 배양되었고 그리고 그들의 혈청은 P. boydii 배양여액 항원과 immunodiffusion test를 실시한 결과 precipitin bands를 형성하였다. 4환자의 흉부 X-선 사진상에는 뚜렷한 진균종음영을 볼 수 없었다. 2명의 환자 객담에서는 P. boydii와 더불어 Candida albicans와 Aspergillus fumigatus가 계속해서 분리 배양되었고 그리고 그들의 혈청에서 그러한 진균들의 항원에 대한 침강항체를 검충할 수 있었다. 3명의 환자는 폐기능이 진균종 제거수술을 허용하지 않았고 1명의 환자는 수술을 거부하였다.

• 대한미생물학회지
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• 제18권1호
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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.

• 한국어병학회지
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• 제1권2호
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• pp.103-110
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• 1988

양식어류의 세균성 질병의 신속한 진단을 위하여 틸라피아(O. niloticus)로부터 분리된 $\beta$용혈성 연쇄구균인 Streptococcus sp. KST-2균주를 사용하여 면역효소법으로 검출실험을 하였으며, 면역효소법의 현장사용의 용이함과 검출의 민감도를 알아보기 위하여 counterimmunoelectrophoresis(CIE), 및 이중면역확산법등과 비교하여 보았다. 다른 어류 병원균과의 교차반응 결과 교차반응이 전혀 일어나지 않는 것으로 보아 이 방법은 $\beta$용혈성 연쇄구균(KST-2균주)에 매우 특이적임을 알 수 있었다. 연쇄구균의 검출실험한 결과 면역효소법은 $1{\times}10^3CFU/ml$의 농도까지 검출가능하였으며, 검출의 민감성이 CIE에 비해 $10^2$배, 이중면역확산법에 비해 $10^4$배나 높은 것으로 나타났다. 면역효소법은 이번 연구에서 사용된 진단법 중에서 현장사용이 가장 용이하며, 민감성이 가장 뛰어난 것으로 나타났다.

• Restorative Dentistry and Endodontics
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• 제8권1호
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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.

• 한국동물학회지
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• 제20권4호
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• pp.169-177
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• 1977

유혈목이(Rhabdophis tigrimus)의 혈장알부민을 polythylene glycol과 ethanol 침전 방법에 의해 정제하였고 혈색소는 agar-gel 전기영동방법에 의해 정제하였다. 이 두 단백질과 유혈목이 혈장을 각각 토끼에 주사하여 항혈청들을 얻었으며 포유류 1종, 조류 1종, 파충류 9종, 및 양서류 1종의 혈장 및 혈구파쇄물과 면역확산 및 면역 전기영동을 실시하였다. 혈장알부민과 혈장에 대한 항혈청들은 유혈목이 혈장알부민과만 반응을 나타냈고 다른종의 혈장알부민과는 반응을 나타내지 않았다. 혈색소에 대한 항혈청은 8종의 사류 혈색소와 강한 반응을 나타냈다. 파충류의 혈장알부민의 구조는 상당히 빠르게 변형되어져 왔고 파충류의 경우 혈색소 분자는 강한 구조상의 상동성을 갖고 있는 것 같다. 그러므로 파충류의 경우 혈색소 분자의 구조는 혈장알부민의 구조보다 느리게 변형되어져 온 것으로 생각된다.

• 한국동물학회지
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• 제32권4호
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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.