• 제목/요약/키워드: indirect-ELISA

검색결과 178건 처리시간 0.031초

서울지역 유기견의 광견병 바이러스 항체 분포 조사 (Serological survey of rabies virus from the stray dogs in Seoul)

  • 김능희;채희선;손홍락;강영일;이정학;김세곤
    • 한국동물위생학회지
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    • 제34권4호
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    • pp.297-301
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    • 2011
  • Rabies virus is transmitted most commonly through a bite from an infected dog. Especially, stray dogs that are not excluded from contact with rabid wild animals can become rabies vectors. Therefore, serological survey of rabies virus from stray dogs in Seoul Metropolitan City was carried out in this study. To investigate prevalence of rabies antibodies in the stray dogs, serum samples were taken from 500 stray dogs between April and December 2010. Antibodies to rabies virus were detected by indirect ELISA. Of 500 tested sera, 147 (29.4%) were positive to rabies virus. Prevalence rates of rabies antibodies(PRRA) in northern and southern Han river region of Seoul were 26.4% and 33.2% respectively. PRRA in male and female dogs were 33.6% and 26.1% respectively. PRRA in less than 1 year, 1~2< years, 2~3< years, 3~5< years, 5~10< years and over 10 years old dogs were 12.7%, 21.6%, 26.4%, 36.4%, 32.5% and 46.4%, respectively. These results indicates that antibody seroprevalence to rabies is still not enough to prevent rabies and rabies vaccination is required to enhance the seroprevalence in the dogs. To improve the situation, measures to raise public awareness of rabies and its prevention is needed. Also, reducing stray dogs and keeping companion animals from contact with wild animals are indispensable for the prevention of rabies.

Production and Characterization of DDT Antibodies and Its Application to Enzyme Immunoassay: Relation of Response and Affinity to Coating Ligand

  • Hong, Ji-Youn;Kim, Jong-Hyun;Park, Song-Ja;Lho, Dong-Seok;Choi, Myung-Ja
    • Bulletin of the Korean Chemical Society
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    • 제24권11호
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    • pp.1605-1608
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    • 2003
  • To development an immunodetection method for DDT, 1,1,1-trichloro-2,2-bis(4-chorophenyl)ethane (p,p'-DDT) and its metabolites (p,p'-DDA, p,p'-DDE, p,p'-DDD), five derivatives of DDT haptens have been synthesised and characterized as coating ligands for antibody evaluation. The appropriate lengths of linkers were introduced to investigate a matching pair of coating ligand and antibody. Among these hapten derivatives, 2,2-bis(4-chlorophenyl)acetic acid (DDA), 5,5-bis(4-chlorophenyl)-5-hydroxypentanoic acid (DDHP) and 5,5-bis(4-chlorophenyl)-5-chloropentanoic acid (DDCP) were conjugated with keyhole limpet hemocyanin (KLH) for its use as an immunogen. The bovine serum albumin (BSA) conjugates of these derivatives were prepared as a coating ligand for monoclonal antibody screening. Fifteen monoclonal antibody clones were screened using these probes. 6,6-Bis(4-chlorophenyl)-6-hydroxyhexanoic acid (DDHH) and 3-[6,6-Bis(4-chlorophenyl)-6-hydroxyhexanoylamino]propanoic acid (DDHHAP), in addition to the above hapten derivatives, were conjugated to ovualbumin (OVA) and bovine serum albumin (BSA) for their use as coating ligands to measure the titration level of the antibody and the displacement of free analytes. The indirect competitive ELISA results indicate that the titration level and free analyte displacement were greatly influenced by the DDT derivatives and carrier proteins used. Three matching pairs of monoclonal antibodies and coating ligands were selected for the DDT immunoassay: antibody clone 1A3 and coating ligand DDA-OVA, 1A1 and DDHHAP-BSA, and 1A4 and DDHP-OVA.

Evaluation of Rhophilin Associated Tail Protein (ROPN1L) in the Human Liver Fluke Opisthorchis viverrini for Diagnostic Approach

  • Geadkaew-Krenc, Amornrat;Grams, Rudi;Phadungsil, Wansika;Chaibangyang, Wanlapa;Kosa, Nanthawat;Adisakwattana, Poom;Dekumyoy, Paron
    • Parasites, Hosts and Diseases
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    • 제58권4호
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    • pp.475-479
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    • 2020
  • Tegumental and excretory-secretory proteins are reported as diagnostic antigens for human opisthorchiasis. Rhophilin associated tail protein1-like (OvROPN1L) protein of Opisthorchis viverrini sperm tail showed potential as a diagnostic antigen. The OvROPN1L recombinant fragments were assayed for diagnostic antigenicity for human opisthorchiasis using indirect ELISA. The strongest antigenic region was a N-terminus peptide of M1 - P56. One synthetic peptide (P1, L3-Q13) of this region showed the highest antigenicity to opisthorchiasis. Sera from other parasitic infections including Strongyloides stercoralis, hookworm, Taenia spp, minute intestinal flukes, Paragonimus spp showed lower reactivity to P1. Peptide P1 is located in the disordered N-terminus of ROPN1L supporting its suitability as linear epitope. In the Platyhelminthes the N-terminal sequence of ROPN1L is diverging with taxonomic distance further suggesting that peptide P1 has potential as diagnostic tool in the genus Opisthorchis/Clonorchis. It should be further evaluated in combination with peptides derived from other O. viverrini antigens to increase its diagnostic power.

Chlamydia trachomatis 진단에 유용한 단세포군 항체 생산에 관한 연구 (Production of Monoclonal Antibody to Chlamydia Trachomatis)

  • 최태열;김신규;김춘원;김기홍;황응수;차창룡;김광혁
    • 대한미생물학회지
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    • 제22권3호
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    • pp.197-208
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    • 1987
  • Chlamydia trachomatis has now shown that this interesting intracellular parasite is a cause of nongonococcal urethritis, infantile pneumonia, pelvic inflammatory disease and epididymitis, in addition to lymphogranuloma venerum and inclusion conjunctivitis. There are several diagnostic methods for C. trachomatis, but the method using monoclonal antibody is the most sensitive and specific. The hybride cell were prepared by fusion of myeloma cell($P_3X_{63}\;Ag_8{\cdot}V_{653}$) of mouse and lymphocyte of mouse(BALB/c) that were immunized with formalin killed C. trachomatis serotype D. The cell mixtures after fusion were dispensed into 640 wells of the 96 well culture plates and continuously cultured in HAT medium for 2 weeks. The supernatants of culture media in 83(13%) wells were reacted with C. trachomatis, which were determined by enzyme-linked immunosorbent assay in 96 well microplate. The clones that secreted antibody to C. trachomatis were cloned by limiting dilution. Only six monoclones secreted antibody to C. trachomatis. The antibody titer of ascitic fluid that collected from same BALB/c mice bearing hybridoma cells was above 1:100,000. These monoclonal antibodies that were IgG reacted with elementary and reticulate bodies of all serotypes(Ba, D, E, F, G, H, J and LGV type-I) using ELISA and indirect immunofluorescence stain, but there were no cross reaction with other bacteria(coagulase negative Staphylococcus, Proteus and E. coli). We concluded these six monoclones secreted the same monoclonal antibody to C. trachomatis. The sensitivity and specificity of the monoclonal antibody compared with Microtrak(confirmatory test of C. trachomatis, Syva) was 100%, respectively.

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SARS-CoV-2 IgG Antibody Seroprevalence in Children from the Amritsar District of Punjab

  • Kaur, Amandeep;Singh, Narinder;Singh, Kanwardeep;Sidhu, Shailpreet Kaur;Kaur, Harleen;Jain, Poonam;Kaur, Manmeet;Jairath, Mohan
    • 대한임상검사과학회지
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    • 제54권3호
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    • pp.173-178
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    • 2022
  • The majority of the children experience milder coronavirus disease 2019 (COVID-19) symptoms. Children represent a significant source of community transmission. Children under 18 years of age account for an estimated 4.8% of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infections globally. However, no conclusive statements pertaining to the multi-fold aspects of the virus in children could be drawn due to the lower prevalence of pediatric cases. The present study was conducted to identify the indirect impact of SARS-CoV-2 infections on developing herd immunity among children in the age group 3 to 18 years by investigating their antibody levels. In the study, 240 children aged 3~18 years were recruited by the Department of Pediatrics, Government Medical College and Hospital, Amritsar, India, and quantification of the antibodies was performed at the Viral Research and Diagnostic Laboratory (VRDL), Government Medical College (GMC), Amritsar, India. Out of the 240 serum samples, 197 (82.08%) showed seropositivity, while 43 (17.92%) were seronegative. When stratified, it was observed that in the age group 3~6 years, 22.33% of children were found to have anti-SARS-CoV-2 antibodies while in the age groups 7~10 years, 11~14 years, and 15~18 years, respectively, 37.06%, 30.46%, and 10.15% were seropositive. Although there was seroconversion among children which was useful for predicting the next wave, no differences in seropositivity were observed between adults and children.

Association of farmers' knowledge, attitude and practices with bovine brucellosis seroprevalence in Myanmar

  • Su Su Hlaing;Satoko Kubota;Kohei Makita;Ye Tun Win;Hnin Thidar Myint;Hiroichi Kono
    • Animal Bioscience
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    • 제37권3호
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    • pp.547-554
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    • 2024
  • Objective: This study aimed to identify the relationship between bovine brucellosis prevalence, farmers' knowledge, attitude, practice (KAP), and social factors on migratory draft cattle and smallholder dairy farms in the central dry zone of Myanmar. Methods: This cross-sectional study was conducted on 54 migratory and 38 dairy cattle farms between August 2020 and February 2021. A structured questionnaire was used to identify farmers' behaviors. Bulk milk was sampled and tested using indirect enzyme-linked immunosorbent assay (I-ELISA). STATA 17 was used for all the analyses. Results: Migratory cattle farms had a higher farm level brucellosis prevalence (14.8%) than dairy farms (2.6%; χ2 = 3.75; df = 1; p = 0.05). Only 2.8% of the farmers had knowledge about brucellosis, while 39.1% and 41.6% had attitudes and farm practices with respect to brucellosis, respectively in the study area. Socio-economic attribute of training in animal husbandry (p<0.01), raising system (p<0.01), practice of separating the aborted cow (p<0.01) were negatively associated to brucellosis. The overall farm level brucellosis prevalence was strongly associated with cattle herd size (p = 0.01), free movement grazing practices (p<0.01), practice of self-removal of placental debris without using personal protective equipment (p<0.01) and farmers' attitudes towards eating cow placenta debris (p<0.01). Conclusion: Farmers had little knowledge of brucellosis. Attitudes and practices differed significantly between migratory and dairy farmers. Training and extension programs are necessary to make farmers aware of their KAP situation since livestock migration and the custom of eating cow placental debris contribute to the spread of brucellosis. Persistent efforts are required to reduce the adverse effects of brucellosis. Therefore, the study suggests that a feasible control intervention and public awareness campaigns need to be conducted regarding methods of preventing human exposure to brucellosis.

가압가열 처리한 시판 돈육 소시지의 항원성에 미치는 소화효소의 영향 (Effect of Digestive Enzymes on the Allergenicity of Autoclaved Market Pork Sausages)

  • 김서진;김꽃봉우리;송유진;이소영;윤소영;이소정;이청조;안동현
    • 한국축산식품학회지
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    • 제29권2호
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    • pp.238-244
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    • 2009
  • 시판 돈육 소시지에 가압가열 처리를 하고 펩신 및 트립신을 처리한 후 ci-ELISA, SDS-PAGE 및 immunoblotting을 실시하여 시판 돈육 소시지의 항원성 변화를 살펴본 결과는 다음과 같다. A와 B 제품 소시지는 가압가열 처리에 의해 항체와의 결합력이 각각 30%, 23% 이하로 크게 감소하였다. SDS-PAGE 결과에서도 두 제품 모두 5분간 가압가열 처리에 의해 PSA band가 약화되었으며 10분 및 30분 처리 시에는 PSA band가 저분자 펩타이드로 많이 분해되었다. Immunoblotting 결과에서는 두 제품 모두 무처리구의 PSA band가 항체와 반응하였지만 가압가열 처리 후에는 항체와 반응하지 않았다. 가압가열 처리에 의한 돈육 소시지의 항원성 저감화를 확인한 후 소화효소가 감소된 시판 소시지의 항원성에 미치는 영향을 살펴보았다. Ci-ELISA를 실시하여 PSA와 p-IgG와의 결합력을 살펴본 결과 A 제품의 경우 5, 10 및 30분 가압가열 처리구가 펩신 및 트립신 처리에 의해 결합력이 약 20% 이하로 크게 감소하였으며 특히 가압가열 30분 처리구는 약 10% 이하의 가장 낮은 결합력을 나타내었다. B제품은 가압가열 30분 처리구의 경우 소화효소 처리 후에도 감소하는 경향을 보였으며 트립신 120분 처리에 의해 약 10% 이하로 결합력이 감소하였다. SDS-PAGE 결과에서는 A 제품의 경우 모든 가압가열 처리구의 PSA band가 펩신 처리 시 저분자 펩타이드로 많이 분해되었으며 10 및 30분 가압가열 처리구의 경우 PSA band가 트립신 처리에 의해 거의 소실되었다. B 제품의 가압가열 처리구는 효소처리에 의해 PSA band가 더욱 약화되었으며 특히 가압가열 30분 처리구의 PSA band는 트립신 처리 후 거의 소실되었다. Immunoblotting결과에서는 A제품의 경우 무처리구의 PSA band가 펩신 30분 처리까지 항체와 다소 강하게 반응하였지만 가압가열과 효소처리 후에는 항체와 반응하지 않았으며 B 제품은 효소처리에 의해 모든 실험구가 항체와 반응하지 않았다. 이상의 결과를 종합해 볼 때 돈육 시판 소시지는 가압가열 처리에 의해 항원성이 크게 감소하였으며 소화효소 처리 후에도 항원성이 증가하지 않고 오히려 감소하는 경향을 나타내었다. 따라서 가압가열 처리한 돈육 소시지는 섭취 후 소화, 흡수 시에도 항원성이 감소하므로 가압가열 처리가 돼지고기 알레르기 저감화에 효과적인 방법임을 확인하였다.

항산화비타민이 Aflatoxin $B_1$ 투여 마우스의 간세포에서 Aflatoxin $B_{1}-DNA$ Adduct 형성에 미치는 영향 (The Effect of Antioxidant Vitamins on Aflatoxin $B_{1}-DNA$ Adduct the Formation in Aflatoxin $B_1$ Administered Mice Liver)

  • 박선자;강성조;박정현;오상석;정덕화
    • 한국식품과학회지
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    • 제33권6호
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    • pp.669-675
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    • 2001
  • 본 연구는 aflatoxin $B_1(AFB_1)$에 의하여 유발된 마우스 간세포에서의 $AFB_1-DNA$ 부가체 형성과 세포의 산화적 손상에 대한 항산화 비타민의 효과를 조사하기 위하여 수행되었다. 먼저, 비타민 C와 비타민 E를 6 주령 수컷 ICR 마우스에 10mg/kg, 63.8mg/kg의 농도로 복강내에 각각 주사(i.p.; intraperitoneal injection) 하였고, $AFB_1$과 비타민 혼합 투여군은 비타민을 주사한 후 1시간이 경과한 다음 0.4mg/kg의 $AFB_1$을 투여하였다. 투여 횟수는 2일 간격으로 동일한 방법에 의해 4회 반복 투여하였다. 반면, $AFB_1$ 단독 투여군은 위와 같은 방법으로 비타민 없이 $AFB_1$만 투여하였다. ELISA에 의한 마우스 혈청내 $AFB_1$의 잔여 농도는 $AFB_1$ 단독 투여군에서 12.28, 18.78 ng/mL이 검출되었다. 그러나 항산화비타민 C 혼합 투여군에서는 7.60 ng/mL, 항산화 비타민 E 혼합 투여군에서는 4.85 ng/mL이 각각 검출되었다. 마우스의간에서 분석된 $AFB_1-DNA$ 부가체 농도에 의하면, $AFB_1$ 단독 투여군에서는 23.78, 25.48 ng/mL이었으며, 항산화 비타민 C 혼합 투여된 군에서는 5.26 ng/mL, 항산화 비타민 E 혼합투여군에서 7.81 ng/mL로 각각 조사되었다. 이들 결과에 대한 통계적 유의성은 p<0.01이었다. 또한 면역조직화학적 관찰에서 $AFB_1$ 단독 투여군에서는 중심정맥과 동양혈관 주변에 갈색의 침전이 두드러지게 나타났으며 이러한 현상은 항산화비타민 혼합 투여군에서 유의하게 감소한 것으로 나타났다.

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전기 검출 시스템을 이용한 Microfluidic Immuno-Sensor Chip (Microfluidic Immuno-Sensor Chip using Electrical Detection System)

  • 맹준호;이병철;조철호;고용준;안유민;조남규;이성환;황승용
    • KSBB Journal
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    • 제21권5호
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    • pp.325-330
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    • 2006
  • Bio-MEMS를 기반으로 microfilter와 백금 전극이 내재되어 있는 microbiochip을 제작하였다. 우리는 이 chip으로 microbead에 indirect ELISA 방법으로 항원-항체를 반응시키고 전기 신호 검출 방법을 이용하여 반응 여부를 판단하였다. 이 때 신호 증폭을 위해 silver enhancer를 사용하였다. Chip 상에서 항원-항체 반응 조건을 최적화하기 위해 pH, temperature, flow time, flow rate, silver enhancer time을 결정하였다. 이렇게 최적화된 조건을 바탕으로 짧은 시간 안에 소량의 시료로 immunoassay를 성공적으로 수행할 수 있었다. 전기 신호 검출 방식을 사용함으로써 biosensor 장비의 소형화와 다중 시료 측정과 자동화를 biosensor에서 적용할 수 있는 가능성을 제시하였다.

Regulation of Chilling Tolerance in Rice Seedlings by Plant Hormones

  • Chu, Chun;Lee, Tse-Min
    • 한국작물학회지
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    • 제37권3호
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    • pp.288-298
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    • 1992
  • Since the major important factors limiting plant growth and crop productivity are environmental stresses, of which low temperature is the most serious. It has been well known that many physiological processes are alterant in response to the environmental stress. With regard to the relationship between plant hormones and the regulation of chilling tolerance in rice seedlings, the major physiological roles of plant hormones: abscisic acid, ethylene and polyamines are evaluated and discussed in this paper. Rice seedlings were grown in culture solution to examine the effect of such plant hormones on physiological characters related to chilling tolerance and also to compare the different responses among tested cultivars. Intact seedlings about 14 day-old were chilled at conditions of 5$^{\circ}C$ and 80% relative humidity for various period. Cis-(+)-ABA content was measured by the indirect ELISA technique. Polyamine content and ethylene production in leaves were determined by means of HPLC and GC respectively. Chilling damage of seedlings was evaluated by electrolyte leakage, TTC viability assay or servival test. Our experiment results described here demonstrated the physiological functions of ABA, ethylene, and polyamines related to the regulation of chilling tolerance in rice seedlings. Levels of cis-(+)-ABA in leaves or xylem sap of rice seedlings increased rapidly in response to 5$^{\circ}C$ treatment. The tolerant cultivars had significant higher level of endogenous ABA than the sensitive ones. The ($\pm$)-ABA pretreatment for 48 h increased the chilling tolerance of the sensitive indica cultivar. One possible function of abscisic acid is the adjustment of plants to avoid chilling-induced water stress. Accumulation of proline and other compatible solutes is assumed to be another factor in the prevention of chilling injuies by abscisic acid. In addition, the expression of ABA-responsive gene is reported in some plants and may be involving in the acclimation to low temperature. Ethylene and its immediate precusor, 1-amincyclopropane-1-carboxylic acid(ACC) increased significantly after 5$^{\circ}C$ treatment. The activity of ACC synthase which converts S-adenosylmethionine (SAM) to ACC enhanced earlier than the increase of ethylene and ACC. Low temperature increased ACC synthase activity, whereas prolonged chilling treatment damaged the conversion of ACC to ethylene. It was shown that application of Ethphon was beneficial to recovering from chilling injury in rice seedlings. However, the physiological functions of chilling-induced ethylene are still unclear. Polyamines are thought to be a potential plant hormone and may be involving in the regulation of chilling response. Results indicated that chilling treatment induced a remarkable increase of polyamines, especially putrescine content in rice seedlings. The relative higher putrescine content was found in chilling-tolerant cultivar and the maximal level of enhanced putrescine in shoot of chilling cultivar(TNG. 67) was about 8 folds of controls at two days after chilling. The accumulation of polyamines may protect membrane structure or buffer ionic imbalance from chilling damage. Stress physiology is a rapidly expanding field. Plant growth regulators that improve tolerance to low temperature may affect stress protein production. The molecular or gene approaches will help us to elucidate the functions of plant hormones related to the regulation of chilling tolerance in plants in the near future.

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