• 약학회지
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• 제45권2호
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• pp.180-189
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• 2001

We have reported that water-extracted Korean mistletoe (KM-110) had various biological activities such as antitumor and immunomodulatory activity, and the pectin fraction (KML-C) of the extract was one of major factors related to its biological functions. In this paper, we produced murine monoclonal antibody (mAb) against KML-C. The cAbs obtained were largely classified into two groups according to specificity to KML-C and ML-I, a pectin from European mistletoe. One group mAbs (9H7-D10 and 3C2-lH4) strongly reacted with KML-C, but not ML-I. In contrast, another group cAbs (8Bll-2C5, BE12-3E9 and 5E10-Fl) reacted with both KML-C and ML-1. The subisotypes of these mobs were shown to be IgGl (9H7-lD10, 3C2-lH4 and 8Bll-2C5) or IgM (8E12-3E9 and 5E10-Fl). To develop an assay system for determination of the amount of KML-C, we established the sandwich ELISA (enzyme-linked immunosorbent assay) method using these mAbs and horse radish peroxidase (HRP)-labelled cAbs. In various combinations of the cAbs for coated antibody and detection antibody, the sandwich ELISA quantitatively detected KML-C, showing the detection limit ranging from 7-5,000 ng/ml. Especially reproducibility (C.V) of the sandwich ELISA, in which 8E12-3E9 was used for coating antibody and 8Bll-2C5-HRP for detection antibody, was 4.59-5.83 in intra assay, and 3.9-9.4 in inter assay.

• 한국동물위생학회지
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• 제22권4호
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• pp.371-375
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• 1999

Total 1,434 sera collected from 72 pig farms were tested for the detection of porcine reproductive and respiratory syndrome (PRRS) virus antibodies. The overall seroprevalence of PRRS virus antibodies was 49.3% (707/727). Of 72 farms tested 59 (81.9%) farms had at least one or more than one pigs with PRRS virus antibodies. The seroprevalence of PRRS virus antibody varied with age. Seroprevalence of PRRS virus antibody in 1 to 30-day-old, 31 to 40-day-old, 41 to 50-day-old, 51 to 60-day-old, and over 61-day-old pig were 27.4%, 52.3%, 57.9%, 52.7%, and 68.2%, respectively. Gilt showed relatively higher seroprevalence (61.2%) than sow (29.2%) and boar (38.3%). In most farms, the infection of PRRS virus was chronic and confined to grower or finisher. This pattern of infection suggests that partial depopulation of the infected herds appears be one of the measures to eradicate the PRRS virus infection. High seroprevalence of the PRRS virus antibody in gilts and boars indicates that the infected gilts and boars in the breeding farms are the major source of the PRRS virus infection, and also play an important role in spreading the PRRS virus between fan mates or herds.

• Parasites, Hosts and Diseases
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• 제25권1호
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• pp.7-12
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• 1987

The indirect fluorescent antibody(IFA) test was used to detect serum IgG and IgM antibodies to Trichomonas vaginalis in 31 vaginal trichomoniasis, 7 candidiasis and in 20 non-infected healthy women with antigen prepared from axonic culture of Trichomenas vaginalis isolated from vulvovaginitis patient. The results were as follows: 1. In 31 vaginal trichomoniasis the positive reactions of IgG antibody were 27 in the 1/8 dilution or higher and :l in the 1/4 dilution whereas in healthy women the reaction showed significantly low as in the 1/4 dilution or below. 2. The sensitivity and specificity of IFA test for IgG antibody to trichomonad antigen in this study were 87.1% and 100%, respectively. 3. No significant difference of IgM antibody levels between vaginal trichomoniasis and healthy women was observed. 4. No relation between the levels of IgG and IsM antibodies to trichomonad antigen by IFA test was observed. 5. No relation between the time lapse and the level of serum IgG antibodies in IFA test of vaginal trichomoniasis was regarded. In conclusion the present study suggests that IFA test in trichomoniasis could be a useful tool for detection of anti-trichomonad IgG antibodies and applicable as an immunodiagnostic method.

• Bulletin of the Korean Chemical Society
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• 제32권6호
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• pp.1975-1979
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• 2011

The modification of a gas diffusion layer (GDL), a vital component in polymer electrolyte fuel cells, is described here for use in the electrochemical detection of antibody-antigen biosensors. Compared to other substrates (gold foil and graphite), mouse anti-rHBsAg monoclonal antibody immobilized on gold-coated GDL (G-GDL) detected analytes of goat anti-mouse IgG antibody-ALP using a relatively low potential (-0.0021 V vs. Ag/AgCl 3 M NaCl), indicating that undesired by-reactions during electrochemical sensing should be avoided with G-GDL. The dependency of the signal against the concentration of analytes was observed, demonstrating the possibility of quantitative electrochemical biosensors based on G-GDL substrates. When a sandwich method was employed, target antigens of rHBsAg with a concentration as low as 500 ng/mL were clearly measured. The detection limit of rHBsAg was significantly improved to 10 ng/mL when higher concentrations of the 4-aminophenylphosphate monosodium salt (APP) acting on substrates were used for generating a redox-active product. Additionally, it was shown that a BSA blocking layer was essential in improving the detection limit in the G-GDL biosensor.

• 대한수의학회지
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• 제45권2호
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• pp.191-197
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• 2005

An immunochromatographic (IC) strip for the rapid detection of Salmonella spp. in the enriched sample was developed. Affinity purified Salmonella polyclonal antibody was conjugated with 40 nm colloidal gold particles which were prepared by citrate method in our laboratory. The antigen-antibody-gold complex was captured by Salmonella antibody attached to test line of nitrocellulose membrane during the capillary migration of sample. Specificity of the IC strip was calculated to be 100% (12/12) and sensitivity was 97.6% (41/42) in the test with pure cultured bacteria. Salmonella was artificially inoculated into raw pork macerated with enrichment broth. And then it was 10-fold diluted from $5.2{\times}10^{8}CFU/ml$ to 5.2 CFU/ml. The IC strip could detect $5.2{\times}10^{6}CFU/ml$ before enrichment. However, the lowest limit of detection was 5.2 CFU/ml after overnight incubation. The results indicated that the IC assay was a rapid, economical and simple method with high specificity and sensitivity for the detection of Salmonella spp. without using any equipment.

• Journal of Biosystems Engineering
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• 제31권2호
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• pp.128-134
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• 2006

Frequent outbreaks of foodborne illness demand the need for rapid and sensitive methods for detection of these pathogens. Recent development of biosensor technology has a great potential to meet the need for rapid and sensitive pathogens detection from foods. An antibody-based fiber-optic biosensor and an automated reagents supply system to detect Listeria monocytogenes were developed. The biosensor for detection of Listeria monocytogenes in PBS and bacteria spiked food samples was evaluated. The automated reagents supply system eliminated cumbersome sample and detection antibody injection procedures that had been done manually. The biosensor could detect $10^4$ cfu/ml of Listeria monocytogenes in PBS. By using the fiber-optic biosensor, $2x10^8$ cfu/ml of Listeria monocytogenes in the food samples were detectable.

• KSBB Journal
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• 제11권4호
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• pp.420-430
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• 1996

열처리된 Salmonella typhimurium 항원을 모델 분석물질로 선택하여 단지 시료만을 첨가함으로써 측정을 수행할 수 있는 1단계 immuno--chromatogra phy 분석시스템이 개발되었다. 개발된 시스템은 분 석물질 특정항체(감지항체)-gold 중합체가 건조된 상태로 축적된 glass fiber membrane(하단), 분석 물질의 다른 epltope를 인지하는 항체(포획항체)와 항 감지항체 특정항체들이 공간적으로 분리되어 고정화된 nitrocellulose membrane(중단) 그리고 흡수대로 선택된 cellulose membrane ( 상단)들로 구성된다. 이와 같은 구성성분들은 분석물질을 포함한 시료수용액이 모세관현상에 의해 연속적으로 이동되도록 부분척으로 포개어 배열되었다. 분석시스템의 성능을 조절하는 변수들은 포획항체의 농도 및 membrane 상의 고정화 위치, membrane 표면처리 와 시료운반용액에 샤용된 비반응성 단백질의 종류, 그리고 중합체의 농도로써 확인되었다. 최적조건하 에서, 시스템의 하부로부터 시료수용액을 흡수 후 15분 이내에 포획항체가 고정화된 membrane 상의 지역에 샌드위치 형상의 항원 항체 면역결합체가 형성되었고 이로부터 발생된 발색신호는 분석물질의 농도에 비례하는 것으로 나타났다. 분석물질의 측정 하한농도는 $1{\times}106$ Salmonella cells/mL인 것으로 나타났다.

• 대한수의학회지
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• 제40권1호
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• pp.81-85
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• 2000

Coxiella burnetii (C burnetii) is the causative agent of Q fever in animal and human. The distribution of the disease has been documented around world. In this study we developed the competitive enzyme linked immunosorbent assay(cELISA) and compared it with indirect immunofluorescent assay(IFA). A monoclonal antibody(Mab) against C burnetii and a peroxidase-conjugated anti-mouse IgM were used as an indicator system competing against antibody in animal serum or as an indicater of the absence of antibody. Sera were considered antibody positive when the percentage inhibition index(PI index) is upper than 30. PI index is calculated as 100-[sample OD/Mab OD)${\times}100$]. Among 162 bovine serum samples, 23 samples were antibody positive both in cELISA and IFA. And 156 samples showed same results. From goat with experimentally induced infection with C burnetii the antibody was detected 20 days early in cELISA compared to IFA. On the basis of present findings, it was demonstrated that cELISA is a reliable diagnostic method for The detection of specific antibodies against C burnetii infection.

• 한국동물위생학회지
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• 제19권1호
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• pp.30-38
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• 1996

Enzyme-linked immunosorbent assay (ELISA) was developed to detect rotavirus antigen in fecal samples using VP6-specific monoclonal antibody(2B12). The ELISA for rotavirus antigen detection found to have specificity to all bovine and porcine rotaviruses tested but not to bovine viral diarrhea virus and bovine coronavirus. The ELISA appeared to have similar sensitivity and specificity compared to fluorescence antibody assay(FA) and electropherotyping (PAGE).

• 대한수의학회지
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• 제10권2호
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• pp.53-57
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• 1970

Hog cholera (HC) virus detection from the artificially infected pigs was made using fluoreescent antibody technique (FAT) and END method. It was observed that the swine origin virulent was detected in most of the organs tested at the early stage of the infection, while the tissue culture attenuated virus was detected only in blood (transitionally), lung, and tonsil.