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Bioluminescence 반응에 의한 ATP 측정을 이용한 젖소 유방염 진단에 관한 연구
김태종,김종배,이승배,전용수,Kim, Tae-jong,Kim, Jong-bae,Lee, Seoung-bae,Jeon, Young-soo 대한수의학회 1989 大韓獸醫學會誌 Vol.29 No.3
This study was carried out to diagnostic test for bovine mastitis by the determination of adenosine triphosphate (ATP) based on firefly bioluminescence. The results obtained are follow; 1. The infection rate of bovine mastitis investigated with 521 cows in 47 dairy farms were found to be 3.6% of clinical form and 44.1% of subclinical form according to the degree of infection. 2. The light yield produced in firefly bioluminescence system was proportional to the concentration of ATP giving stright line within the range of 100PM~1uM. 3. When the number of somatic cell in milk was determined by the ATP assay and compared with three conventional methods such Fossomatic, California mastatic test (CMT), and rolling ball viscometer (RBV), it was shown that r=0.92 for Fossomatic, 0.63 for CMT and 0.7 for RBV. 4. The microorganisms causing mastitis were isolated Staphylococcus sp. (53.3%), Streptococcus sp. (17.9%), Micrococcus sp. (13.5%), Gram negative bacilli (6.3%), Gram positive bacilli (5.5%) and Yeast-like fungi (5.4%). 5. The endogeneous ATP levels of bacteria in a raw milk determined by the firefly bioluminescence system and compared with the results of the conventional methods. The correlation was 0.88 for raw milk.
Bioluminescence 방법에 의한 우유 및 유제품에 존재하는 세균수 측정에 관한 연구
이승배,도형기 건국대학교 1986 論文集 Vol.22 No.1
Determination of bacterial ATP based on luciferin-luciferase bioluminescence reaction was applied to the measurement of bacteria in raw milk and liquid yoghurt as in alternative rapid method. The results are as follows : 1. The light yield was proportional to the concentration of ATP giving a straight light within a range of 10-10 to 10-6M. 2. The light yield related with the ATP content was increased as the number or bacteria in E. coli and L. bulgaricus was increased. When ATP content of bacteria were determined by bioluminescence reaction and compared. 3. The patterns of growth curve for E. coli look very similar, when they were monitored by ATP content assayed by bioluminescence reaction and total colony count. 4. The bacterial ATP of each sample in raw milk and liquid yoghurt was proportional to the number of bacteria, Coefficient of correction was 0.74 and 0.58 respectively. Thus, this new rapid method based on determination of bacterial ATP monitored by luciferin-luciferase bioluminescence reaction can be used for the indirect counting of bacteria, respectively.
Staphyloccus aureus에 대해 gentamicin의 MIC측정을 위한 firefly Iuciferase assay의 응용
이승배,최상훈,이동만 건국대학교 1987 論文集 Vol.24 No.1
Susceptibllity of Staphyloccus aureus (ATCC 14492) to gentamicine was determined by minium inhibitory concentration(MIC) test and firefly bioluminescence assay of bacterial adenosine triphosphate(ATP). ATP content extracted Staphyloccus aureus was proportional to the number of bacterial, ATP content in a bacteria was 3.1fmol. The patterns of growth curves for Staphyloccus aureus look very similar, when the bacterial growth was monitored by ATP content and viable cell count. The MIC of gentamicine for Staphyloccus aureus measured both firefly bioluminescence assay and MIC test was 0.5ug. Determinent of intrcellular ATP in culture of Staphyloccus aureus exposed to gentamicin seem to be suitable for reliable determination of MIC within 3 hours of incubation. It gave very good correlation(r=0.97), when firefly bioluminescence and MIC test were compared.
Bacillus thuringiensis 아종 Kustaki의 내독소 결정체에 대한 항체생산과 항혈철중 면역글로부린 G의 분리
이승배,김창규 건국대학교 1986 論文集 Vol.23 No.1
The antisera were obtained by inoculating rabbits with purified solublized crystal toxins from Bacillus thuringiensis subspp. Kustaki HD1 and HD73. Enzyme-linked Immunosorbent Assay(ELISA) was used to detect anti crystal toxin antibody. Bacillus thuringiensis subspp. kutaki and HD73 crystal toxins were purified by Renografin-76 density gradient centrifugation. The density of crystal toxin was 1.22g/cm3. When crystal toxins were infected into rabbits for producing anti-crystal toxin antibody, the good results were obtained at 4 or 5 times injections with 50 to 200og crystal toxin. Immunoglobulin G in antisera was purified by Protein-A Affinity Chromatography. The heavy chain and the light chain of immunoglobulin G were seperated by SDS-polyacrylamide gel electrophoresis. Then molecular weights of heavy chain and light chain were approximately 50,000 daltons and 25,000 daltons.
hCG에 대한 단일클론 항체 생산과 그 이용에 관한 연구
崔尙勳,李丙哲,李承培 건국대학교 1987 論文集 Vol.25 No.1
This study was carried out to produce monoclonal antibody against hCG secreted by the tr-ophoblast of developing embryo. Produced monoclonal antibody to hCG was regarded as IgG class since it had a strong affinity for a protein A. The concentration of monoclonal antibody in cell culture supernatant and in mouse ascitic fluid was 250㎍ and 30mg perml, respectively. The titer of antibody in mouse ascitic fluid was about 1 : 106~107 on ELISA
Choi, Sang-Hoon,Lee, Seoung-Bae,Lee, Hoon-Bhum 건국대학교 1987 論文集 Vol.24 No.1
α-Fetoprotein 측정을 위한 chemiluminoimmunometric assay는 AFP내의 서로다른 위치에 존재하는 항원 결정기를 인지하는 두개의 단일클론 항체를 이용한 분석방법으로서 polystyrene tube에 두가지의 항체중 한가지를 coating 시키고 그위에 unknown 또는 known AFP을 반응시킨후 추적물질로서 화학발광체를 합성한 또 다른 항체를 반응시켜 최종 detection은 Luminometer(LKB, 1250)를 이용 측정하였다. 그 결과 감도가 상당히 좋은 표준곡선을 얻을 수 있었으며 기존의 방법(Radioimmunometric assay)과 비교해 볼때 더 나은 detection limit를 얻을 수 있었다. 이러한 신속하고도 정확한 방법을 임상진단에 응용하면 상당한 잇점이 있으리라 사료된다.
생물학적 발광반응에 의한 액상발효유 내의 유산균 수 측정에 관한 연구
韓錫絃,金鍾培,李承培 건국대학교 1988 새마을연구논문집 Vol.7 No.-
Determination of bacterial ATP based on luciferin-luciferase bioluminescence reaction was applied to the measurement of bacteria in liquid yoghurt as an alternative rapid method. The results are as follows; 1. When the number of bacteria such as Streptococcus thermophilus, Lactobacillus, acidophilus, Lactobacillus bulgaricus were counted by conventional agar plating technique and compared with the concentration of bacterial ATP, it gave a good linear relationship. The content of ATP per bacteria cell was 3.2∼4.6 fM depending on the kinds of lactic acid bacteria. 2. In order to investigate the most efficient method to remove the non-bacterial ATP, the various methods of hydrolyzing with ATPase, filtration and centrifugation were compared. The hydrolyzing method was found to be the most efficient in terms of reproducibility, speed and simplicity. 3. The endogeneous ATP levels of Lactic acid bacteria in liquid yoghurt was determined by the firefly bioluminescence system and compared with the result of conventional agar plating technique. The correlation coefficient was 0.83 between two methods. 4. This new rapid method (<lhr.) bases on the determination of bacterial ATP monitored by luciferin-luciferase bioluminescence reaction can be for the indirect counting of Lactic acid bacteria in liquid yoghugrt.