• 한국미생물·생명공학회지
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• 제20권2호
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• pp.225-232
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• 1992

효소면역측정법에 의한 aflatoxin $B_1(AFB_1)$의 정량법을 개발하기 위하여, 항체를 생산.정제하여 분석법을 확립하고 직접법과 간접법(direct/indirect competitive ELISA)의 특성 및 문제점을 비교. 검토하였다. Bovine serum albumin(BSA)을 carrier protein으로 한 $AFB_1$-1-(O-carboxymethyl)oxime -BSA를 토끼에 면역하여 항$AFB_1$항혈청을 생산하였다. 정량 침강반응에 의하여 항혈청으로부터 항BSA항체를 제거하고 황산암모늄 침전법 및 EDAE-Sephadex A-50 이온교환 크로마토그래피를 통하여 순도 높은 IgG항체를 정제하여 항$AFB_1$항체를 사용하였다.

• 한국동물위생학회지
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• 제26권3호
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• pp.215-219
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• 2003

This study was conducted to investigate the similarity between hemagglutination inhibition (HI) test and enzyme-linked immunosorbent assay(ELISA) titers and sample to positive ratio (S/P ratio) of Newcastle disease(ND) virus. To perform this study, the 372 sera of broiler chicks and 120 sera of layers and breed chicks were collected from slaughter house and farms, respectively. As a result of HI test out of different chicks, the positive percentage of ND antibody titer of broiler, layer and breeder, when a standard positive HI titer were '2', was 84.4%, 100% and 100%, respectively. The positive percentage of ND antibody titer by ELISA was shown 38.4%, 100% and 100% and S/P ratio were also shown 81.5%, 98.2% and 99.2%, respectively. The results of comparative survey with same sera by two experimental methods were as follows; In low HI titer, ELISA titer was not similar to HI titer, but S/P ratio was similar to it. In high HI titer, ELISA titer was not similar to HI titer. Therefore, HI titer was more similar to S/P ratio than ELISA titer.

• 대한수의학회지
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• 제54권3호
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• pp.147-150
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• 2014

Q-fever is a vector-borne (Coxiella [C.] burnetii) zoonotic disease that is an increasing public health concern. To date, some research about Q-fever prevalence in dairy herds and human patients has been reported in Korea, but information about Korean native cattle is scarce. To measure the prevalence rates of C. burnetii in Korean native cattle, a total of 1,095 bovine serum samples collected during 2010~2013 were analyzed with an enzyme-linked immunosorbent assay. Sixty-eight heads of cattle were diagnosed as positive and while 19 heads were suspected (positive rate = 6.2%). Interestingly, Jeju province had a seropositivity rate six times greater than that of other provinces (18.9% vs. 3.2%). High seroprevalence might be caused by wide distribution of ticks in Jeju province compared to other regions. Based on these data, extensive monitoring of C. burnetii infection in cattle, tick distribution, and climate changes is required.

• 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.

• 한국식품위생안전성학회지
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• 제13권2호
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• pp.129-134
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• 1998

Nivalenol(NIV)의 검출을 위한 효소면역측정법(ELISA)을 개발하기 위하여 tetraacetyl nivalenol(Ac4-NIV)에 대한 다클론항체를 생산하고 그 조건을 확립하였다. Ac4-NIV-hemisuccinate를 bovine serum albumin에 공유결합 시킨 Ac4-NIV-HS-BSA를 Freund's adjuvant와 함께 수차례 토끼에 피하면역하였다. 가장 높은 항체가를 나타낸 항혈청으로부터 정제한 항체와 Ac4-NIV-HS-HRP conjugate를 이용하여 직접 경합 ELISA(cdELISA)를 확립하였다. 그 표준 곡선으로부터 Ac4-NIV의 검출 범위는 10~5,000 ng/ml(ppb)임을 알 수 있었다. 특이항체의 Ac4-NIV과 acetyl T-2에 대한 반응성은 각각 100, 70%였으나, NIV, deoxynivalenol, 3-acetyl deoxynivalenol, 15-acetyl deoxynivalenol, triacetyl deoxynivalenol, fusarenon-X, T-2에 대한 반응성은 0.1% 이하로 극히 미약하였다. NIV를 인위적으로 오염시킨 옥수수 시료를 70% acetonitrile 추출하고 acetylation 한 다음 cdELISA를 행하였을 때, 분석의 회수율은 100, 300, 1,000 ng/g(ppb)에서 각각 108, 143, and 70%(평균, 107%)로 나타났다.

• 한국식품과학회지
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• 제32권3호
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• pp.720-725
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• 2000

여러 가지 처리가 난백의 항원성을 감소시키는데 미치는 영향을 조사하기 위하여, 토끼 항난백항체로 난백 처리물들의 간접 경합 ELISA(ciELISA)를 실시하였다. 식물성, 동물성, 미생물성의 9가지 효소를 이용한 가수분해처리는 난백의 항원성에 영향을 주지 못했다. 보다 효율적인 효소 가수분해를 위해 방사선 조사 후, 효소처리를 실시했는데, 이것 역시 항원성을 감소시키지 못했다. Trifluoromethanesulfonic acid 처리는 난백의 항원성을 거의 제거시키는 것으로 나타났다. 열처리는 $121^{\circ}C$ 고온처리의 경우에서만 반응성이 1/10,000이하로 감소하여 항원성이 거의 사라졌다. NaOH 단독 처리는 0.3%(w/v) 이상의 농도에서 반응성이 감소되기 시작하여, 1%(w/v) 이상에서는 반응성이 1/10,000이하로 감소하였다. NaOH 처리에 $70^{\circ}C$, 15분간의 열처리를 더해 주면, NaOH 단독 처리시보다 효과적으로 항원성이 감소되는 것으로 나타났다.

• 한국임상수의학회지
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• 제27권4호
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• pp.402-406
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• 2010

Bovine leukemia virus (BLV) is a delta-retrovirus which causes chronic lymphocytosis in cattle. BLV infections have been divided into two groups such as enzootic bovine leukosis (EBL) and sporadic bovine leukosis (SBL) according to the clinical symptoms in infected cattle. The conventional detection method of BLV was hematological procedure which is determining lymphocytosis in the suspected animals. Recently several sensitive methods were developed to detect antibody to BLV and nucleic acid of the BLV from infected cattle. In this study we have compared the difference of positive rates between agar gel immunodiffusion (AGID) and enzyme linked immunosorbent assay (ELISA) which are using for BLV antibody detection methods. The positive detection rate of ELISA test was 7.4% greater than the positive rate of AGID. The discrepancy of the positive rate between ELISA and AGID were showed in the group of age over one year old to under three year old group. The result from each test agreed very well in the group of over 5 year old cattles. The serological test is very useful method to select the infected cattle for the eradication or control of the disease in the infected herd. But it has a limit by interference of the maternal antibody from the cow of under 6 month old. This study shows that 16.2% of these ages group showed BLV gene positive by polymerase chain reaction (PCR) method. The result suggests that ELISA test need to be used with PCR to clarify misinterpretation of positive animals by antibody response due to the natural infection from maternally derived antibody in calves of under 6 months old.

• 한국식품영양과학회지
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• 제29권6호
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• pp.1030-1034
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• 2000

계란의 방사선 조사 여부를 효소면역 측정법에 의해 판별 가능한지를 검토하기 위해, 방사선에 대해 가장 민감한 것으로 알려진 오브알부민의 항 오브알부민 19G에 대한 반응성을 경합효소면역측정법 (cELISA)에 의해 검토하였다. 계란은 난각이 있는 상태에서 조사선량은 0~7 kGy로 조사하였다. 항 오브알부민 IgG에 대한 오브알부민의 반응성은 조사선량이 증가함에 따라 감소하였는데, 감소 정도는 조사선량 의존적으로 변화되어 7 kGy에서 가장 크게 감소하였다. 방사선 조사에 의한 반응성감소 정도는 1.5배 (0.5 kGy)~3.7배 (7 kGy)이었다. 가장 조사선량이 낮은 0.5 kGy에서도 반응성의 감소가 측정 가능했기 때문에 cELISA는 저선량 조사된 계란의 검출에 유용한 방법으로 사용될 수 있을 것으로 사료된다. 방사선 조사에 의한 오브알부민의 반응성 감소는 오브알부민의 부분적 분해에 의해 구조적 변화가 일어났기 때문으로 사료된다.

• The Plant Pathology Journal
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• 제17권1호
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• pp.62-66
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• 2001

Detection of Xanthomonas axonopodis pv. citri (Xac) on citrus fruits for exporting is usually made by bacteriophage test (BPT) to demonstrate the pathogen-free status. BPT has rather time-consuming and complicate procedures for dealing with massive samples to be inspected. In this study, enzyme-linked immunosorbent assay (ELISA) was applied to detect Xac on fruits, and compared with BPT. In ELISA, positive reactions occurred in the bacterial densities of $3\times10^5$ cells/ml or more. To detect the bacterial infection on citrus fruits with a density of lower than $3\times10^5$ cells/ml, the bacterial suspensions were mixed with fruit rinse water and incubated in broth medium. Ordinary peptone sucrose broth (PSB) was not a proper medium for increasing Xac density specifically enough to be detect by ELISA. On the other hand, modified PSB (MPSP) amended with Fe-EDTA (0.25 g/$\ell$) and 2.5% potato-dextrose broth sufficed to differentiate uninfected and infected citrus fruits by ELISA after 24 h incubation of the fruit rinse water. Using various citrus samples from infected and uninfected fields, efficiencies in detecting Xac on fruits were compared between ELISA and BPT. For infected fruits samples, ELISA detected Xac by 100%, while BPT by about 44%, indicating that the detection efficiency was improved by 23.5% by ELISA, compared to BPT. In addition, ELISA has simpler procedures for testing and is less time-consuming than BPT, suggesting that ELISA may be accurate and simple method to detect Xac on citrus fruits.

• 한국식품과학회지
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• 제32권6호
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• pp.1221-1226
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• 2000

대두단백질의 분석을 위한 효소면역측정법을 개발하고자 특이항체를 생산하고 그 항체의 특성을 비교하였다. 분리대두단백(ISP), ISP를 SDS와 urea의 첨가하에 열처리한 것(ISP(SU)), 11S 글로불린의 acidic subunit(AS)를 각기 면역하여 다클론 항체를 생산하였다. 간접 경합 효소면역측정법(ciELISA)을 실시하여 처리를 달리한 대두단백질에 대한 이들 항체의 반응성을 조사하였여 $IC_{50}$으로 나타내었다. 항AS 항체의 경우 $IC_{50}$값이 ISP, ISP(SU), ISP를 2-ME로 처리한 ISP(ME), crude 11S에 대하여 각각 20, 2, 2.5, $200\;{\mu}g/mL$로 나타났다. 또한, 항ISP(SU) 항체의 경우 같은 항원에 대해서 100, 5, 4, $220\;{\mu}g/mL$였으며 항ISP 항체의 경우는 각각 20, 30, 36, $1000\;{\mu}g/mL$로 나타났다. 이로서 생산된 3가지 항체 중에서 항AS 항체가 대두단백질에 대한 반응성이 가장 우수하여 ELISA분석법 개발에 적합하였다.