• Plant Biotechnology Reports
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• 제2권2호
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• pp.137-143
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• 2008

Production of Indian citrus ringspot virus (ICRSV)-free plants from an infected plant of kinnow mandarin (Citrus nobilis Lour ${\times}$ C. deliciosa Tenora) is reported. The shoot apices of different sizes (0.2-1.0 mm) excised from the ICRSV-infected plant were micrografted onto decapitated rootstock seedlings of rough lemon (C. jambhiri). Micrograft survival depended on the size of shoot apex and the sucrose concentration of the culture medium. Increase in scion size from 0.2 to 0.7 mm resulted in an increase in micrografting success rate from 30.55 to 51.88%. Further, micrograft survival obtained with 0.2 mm was improved from 30.55 to 38.88% by increasing sucrose concentration in the culture media from 5 to 7.5%. The micrografted plants were tested for ICRSV using ELISA and RT-PCR. All plants raised from 0.2-mm scion were found negative with both ELISA and RT-PCR whereas only 20% of the ELISA negative plants raised from 0.3-mm scion were found negative for ICRSV with RT-PCR. The outcome of this research is the successful establishment, acclimatization and virus testing of micrografted plants.

• 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.

• 한국동물위생학회지
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• 제39권4호
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• pp.239-246
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• 2016

The prevalence of Coxiella burnetii in native Korean goats in Jeonbuk province was investigated using ELISA (sera) and PCR (vaginal mucus). A total of 798 blood samples from 189 farms were collected in 2013 and 2015. Seroprevalence was 13.6% in 2013 and 15.7% in 2015. Tracking survey of six seropositive farms on the prevalence of C. burnetii was conducted. 137 (41.4%) out of 331 goats were seropositive and 47 (17.6%) out of 267 goats were positive in PCR. The higher seropositivity observed in adult goats and female goats significantly, the seroprevalence of C. burnetii was higher as age increased (<1 y=13.2%~${\geq}5$ y=100%)(P<0.05) and female goats (44.8%) was higher than male goats (24.1%) on the seroprevalence (P<0.05). 21 (7.9%) goats positive in PCR was seronegative. The prevalence of C. burnetii in native Korean goats in slaughter house was 30.0% in ELISA and 11.5% in PCR. Significantly, female goats (62.8%) was higher than male goats (12.3%) on the seroprevalence (P<0.05). Based on these data, C. burnetii was easily exposed in domestic animals: native Korean goats-related areas such as farms and slaughter house.

• Journal of Veterinary Science
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• 제22권4호
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• pp.45.1-45.13
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• 2021

Background: Peste des petits ruminants (PPR) is an infectious disease caused by the peste des petits ruminants virus (PPRV) that mainly produces respiratory symptoms in affected animals, resulting in great losses in the world's agriculture industry every year. Single-domain variable heavy chain (VHH) antibody fragments, also referred to as nanobodies, have high expression yields and other advantages including ease of purification and high solubility. Objectives: The purpose of this study is to obtain a single-domain antibody with good reactivity and high specificity against PPRV. Methods: A VHH cDNA library was established by immunizing camels with PPRV vaccine, and the capacity and diversity of the library were examined. Four PPRV VHHs were selected, and the biological activity and antigen-binding capacity of the four VHHs were identified by western blot, indirect immunofluorescence, and enzyme-linked immunosorbent assay (ELISA) analyses. ELISA was used to identify whether the four VHHs were specific for PPRV, and VHH neutralization tests were carried out. ELISA and western blot analyses were used to identify which PPRV protein was targeted by VHH2. Results: The PPRV cDNA library was constructed successfully. The library capacity was greater than 2.0 × 10⁶ cfu/mL, and the inserted fragment size was approximately 400 bp to 2000 bp. The average length of the cDNA library fragment was about 1000 bp, and the recombination rate was approximately 100%. Four single-domain antibody sequences were selected, and proteins expressed in the supernatant were obtained. The four VHHs were shown to have biological activity, close affinity to PPRV, and no cross-reaction with common sheep diseases. All four VHHs had neutralization activity, and VHH2 was specific to the PPRV M protein. Conclusions: The results of this preliminary research of PPRV VHHs showed that four screened VHH antibodies could be useful in future applications. This study provided new materials for inclusion in PPRV research.

• 한국식품과학회지
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• 제35권6호
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• pp.1209-1215
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• 2003

가시오가피 냉침 추출물에서 면역자극활성을 가지는 지표물질을 분리하고자, gel chromatography를 수행하여 비장세포에 대한 증식활성을 가지는 EN-SP을 분리하였고, 생화학적 분석 결과 단백 다당 물질임을 확인하였다. 가시오가피 추출물 내에서 지표물질로서 EN-SP 성분을 정량하기 위하여 경쟁적 ELISA법을 개발하여 다음과 같은 결과를 얻었다. 경쟁적 ELISA법을 이용하여 EN-SP-HRP conjugate의 희석농도를 1000배로 결정하였으며, EN-SP의 표준곡선을 작성한 결과 검출범위는 $0.2{\sim}20\;{\mu}g/mL$으로서 측정감도는 $0.2\;{\mu}g/mL$이었다. 이러한 표준곡선의 정당성 평가를 위하여 정량된 EN-SP를 통한 재현성 실험에서 $5\;{\mu}g/mL$ 이하의 농도에서 intra assay 경우 C.V. 값은 $6.13{\sim}8.81%$, bias는 평균 -3.7%를 보였으며, inter assay에서도 C.V. 값과 bias 모두에서 10%이내로 비교적 우수한 재현성을 보였다. 작성된 표준곡선을 바탕으로 유사물질인 오가피의 $4^{\circ}C$ 추출물들의 EN-SP의 함유량을 조사한 결과 EN-SP 성분은 주로 가시오가피 외피에서만 높은 함유량을 보였다. 동일한 추출물의 macrophage 자극에 의한 cytokine 유도활성 실험결과, EN-SP 성분을 함유하는 가시오가피의 수피 추출물만 IL-12 및 $TNF-{\alpha}$를 생산하는 면역자극활성을 나타냄으로서 EN-SP 성분은 가시오가피에서 면역자극활성을 나타내는 지표물질로의 가능성이 제시되었다.

• Parasites, Hosts and Diseases
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• 제56권5호
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• pp.437-446
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• 2018

To investigate the prevalence of Toxoplasma gondii in pork on the market in Korea, an in-house enzyme-linked immunosorbent assay for tissue fluid (CAU-tf-ELISA) was developed using a soluble extract of T. gondii RH strain tachyzoites. As the standard positive controls, the piglets were experimentally infected with T. gondii: Group A (1,000 cysts-containing bradyzoites), Group B (500 cysts-containing bradyzoites) and Group C ($1.0{\times}10^3$ or $1.0{\times}10^4$ tachyzoites). The CAU-tf-ELISA demonstrated infection intensity-dependent positivity toward tissue fluids with average cut-off value 0.15: 100% for Group A, 93.8% for Group B and 40.6% for Group C. When tissue-specific cut-off values 0.066-0.199 were applied, CAU-tf-ELISA showed 96.7% sensitivity, 100% specificity, 100% positive and 90.0% negative predictive values. When compared with the same tissue fluids, performance of CAU-tf-ELISA was better than that of a commercial ELISA kit. Of the 583 Korea domestic pork samples tested, anti-T. gondii antibodies were detected from 9.1% of whole samples and 37.9% from skirt meat highest among pork parts. In the 386 imported frozen pork samples, 1.8% (skirt meat and shoulder blade) were positive for anti-T. gondii antibodies. In Korea, prevalence of anti-T. gondii antibodies in the pork on retail markets appeared high, suggesting that regulations on pig farming and facilities are necessary to supply safe pork on the tables.

• 한국식품위생안전성학회지
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• 제3권2호
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• pp.65-73
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• 1988

종래의 화학적, 생물학적 분석방법의 단점인 검출한도가 낮은점, 시료전처리의 복잡성, 비경제성 및 비 특이성 등을 극복하기 위해 monoclonal AB를 사용한 ELISA법으로 T-2 toxin 에 대해 특이성있는 새로운 분석법을 개발하여 다음과 같은 결과를 얻었다. 1. 종래의 GLC 및 GC-MS 분석법보다 간편하고 신뢰성이 높으며 검출한계가 0.1ppb인 고감도의 분석법을 개발하였다. 2. 본 분석법을 이용하여 Fusarium spp. 균의 T-2 생산유무를 단시간에 다량의 시료를 검색할 수 있었으며, data의 정확도가 GLC와 유사하며 GLC로는 검색할 수 없는 150ppb이하의 미량함유 시료에서도 T-2 toxin을 검색할 수 있었다. 3. 본 실험에 사용한 F. sporotichioides M-1-1 균주의 밀에 대한 최적 배양조건은 $24~27^{\circ}C$ 2주간임을 알았다.

• 한국식물병리학회지
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• 제3권4호
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• pp.261-269
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• 1987

포장에서 고추에 황화, 엽맥록대 그리고 엽맥주름을 일으키는 바이러스가 분리되었다. 이 바이러스는 PVY항혈청과 ELISA 검정에서 반응하였으나 CMV, TMV, TBRV, AMV, TSWV, TEV, PMV, TRSV와는 반응하지 않았다. 바이러스 입자의 전자현미경 검경 결과 길이가 750-760nm의 사상형 입자였다. 이 바이러스는 진딧물에 의해 쉽게 전염되었으며 기주범위는 PVY와 비슷하였으나 Chemopodium amaranticolor와 C. quinoa에 전신 감염을 일으켰다.

• Toxicological Research
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• 제19권2호
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• pp.133-139
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• 2003

Allergic contact dermatitis may be caused by a wide variety of chemicals. A murine local lymph node assay (LLNA) has been developed as an alternative to guinea pig models for assessing the contact sensitization potential of chemical. However, there is a need to develop a nonradioisotopic endpoint for the LLNA, because of the radioisotopic method's requiring the use of special facilities. In this study, we investigated the development of a nonradioisotopic endpoint for LLNA using ELISA (enzyme-linked immunosorbent assay). Female Balb/c mice were treated by the topical application on the dorsum of both ears with four different strong sensitizers, 2,4-dinitrochlorobenzene (DNCB), oxazolone (OXZ), toluene diisocyanate (TDI), and trimellitic anhydride (TMA), and a strong irritant, sodium lauryl sulfate (SLS), once daily for three consecutive days. The proliferation of cells in the auricular Iymph node was analyzed by means of the labelling index (Ll) of bromodeoxyuridine (BrdU) incorporation into cells. The weights of the Iymph nodes in the mice treated with allergens, DNCB, OXZ, TDl and TMA were increased compared to the vehicle control. The stimulation index (Sl) of mice treated with DNCB, OXZ, TDl, and TMA was over three-fold increase compared to the vehicle control. However, the S1 of mice exposed to SLS was not significantly increased compared to the vehicle control, while the lymph node weight of SLS was significantly increased. These results suggest that the LLNA modified endpoint using ELISA based on BrdU incorporation could provide a useful method of screening for irritants and allergens.

• 대한수의학회지
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• 제40권1호
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• pp.130-137
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• 2000

In order to diagnose canine heartworm infection by antigen capture ELISA, the crude somatic(S), partial somatic(below 45kDa) and excretory/secretory(E/S) antigen of adult heartworm were identified and the antigenicity was examined by silver stain, immunoblot and ELISA. Then, production of monoclonal antibody to specific antigen carried out in this experiment. The bands to S antigen and E/S antigen were recognized between 10 and 200kDa and common bands were recognized strongly 14, 18, 28, 43kDa by silver stain. By western blot analysis, fractions to S antigen were recognized 14, 16, 18, 20, 24, 28, 32, 43, 50, 55kDa, etc. and only a 14kDa to E/S antigen in positive sera which were positive in modified Knott's test and necropsy. In ELISA, the positive sera reacted to antigens(SA, $SA_{45}$, E/S) were significantly different from negative sera by Student's t-test(p<0.05). Four hybridoma cell lines(14, 16, 17, 32kDa) than produce specific monoclonal antibodies for these antigens were obtained by immunizing BALB/c mice with a partially purified somatic antigen (below 45kDa) preparation, by fusing spleen cells with SP2/O cell myeloma cells, and by screening cell culture supernatants for antibody. In these results, it was confirmed that partial somatic antigen(below 45kDa) or E/S antigen can be used for serologic diagnosis of heartworm infection and monoclonal antibody reacting with specific antigen(14kDa) can be used for antigen capture ELISA in prepatent period of canine heartworm infection.