• 제목/요약/키워드: ELISA-inhibition test

검색결과 49건 처리시간 0.029초

배양조건이 Aspergillus flavus ATCC 15517의 Aflatoxin 생성에 미치는 영향 (The Effect of Cultural Conditions on the Aflatoxin Production of Aspergillus Flavus ATCC 15517)

  • 정덕화;이용욱;김용호;김성영;김종규
    • 한국환경보건학회지
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    • 제16권1호
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    • pp.45-53
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    • 1990
  • To investigate the effect of cultural condition on the aflatoxin production of Aspergillus flayus ATCC 15517, mixed culture with Aspergillus niger, better kind of media and size of Cultural vessels were examined. YES medium was better than SLS medium for this study. Small scale test tube culture was showed the possibility to simply examine the growth, total acidity, pH and aflatoxin production during cultivation, and also could reduce the second contamination of aflatoxin B1 from large scale broth cultured. Especially ELISA method is simple, sensitive and specific and therefore well suited to small scale of test tube culture. Mixed culture significantly reduced the aflatoxin production of Aspergillus fiavus ATCC 15517 and showed almost 95% inhibition of that level during the incubtation.

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Serologic monitoring of animal welfare-oriented laying-hen farms in South Korea

  • So, Hyunhee;Jeong, Seolryung;Mo, Jongsuk;Min, Kyungchul;Kim, Jongnyeo;Mo, In-Pil
    • 대한수의학회지
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    • 제58권4호
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    • pp.193-199
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    • 2018
  • As animal welfare issue becomes important, the European Union bans conventional cages for laying hens from 2012. So the alternative housing systems like floor pens, aviaries or free range systems have been suggested. From 2011 to 2014, we monitored 20 welfare-oriented laying hen farms in South Korea to figure out serological status of major viral diseases. During this period, total 3,219 blood samples were collected from the randomly selected chickens to test and evaluate the hemagglutination inhibition titers for low pathogenic avian influenza, Newcastle disease and egg drop syndrome '76. A total of 2,926 blood samples were tested through enzyme linked immunosorbent assay (ELISA) to assess the serological status of infectious bronchitis (IB). The distribution of ELISA titers for IB was various from almost 0 to 20,000 through the all weeks of age. Also, the antibody coefficient of variation for most of the diseases in this study was higher than those of typical cage layers. As this study was the first surveillance for major avian viral diseases of the animal welfare-oriented farms in South Korea, the results obtained from this study will help to determine what information and resources are needed to maintain better biosecurity and to improve the health and welfare of laying hen flocks.

우유로부터 Osteopontin의 분리.정제 및 특성에 관한 연구 (Purification and Properties of Osteopontin from Bovine Milk)

  • 최기원;김동운;이수원
    • Journal of Animal Science and Technology
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    • 제45권3호
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    • pp.491-498
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    • 2003
  • 본 연구는 우유로부터 OPN을 분리 정제하여 그 특성을 규명하기 위해서 수행되었다. 먼저 ion-exchange와 hydrophobic chromatography를 이용하여 우유로부터 OPN을 분리 정제하였다. OPN의 분자량은 SDS-전기영동 상에서 약 60,000dalton이었고, NH2-terminal amino acid sequence를 확인한 결과 Leu-Pro-Val-Lys-Pro- Thr-Ser 순이었다. OPN을 35주령된 SCWL를 이용하여 산란계를 면역 시키고, 형성된 anti- OPN IgY 항체를 분리․정제한 후 ELISA test로 항체가를 측정하였다. 또한 RID test을 이용하여 OPN 함량에 따른 정량곡선을 작성하고, 이 곡선에 의해 우유 중 OPN 함량을 정량하였다. 그 결과 원유, 탈지유, 시유에서 각각39.78, 31.74, 37.48${\mu}g$/$m\ell$을 함유하고 있는 것으로 나타났다. 또한 OPN의 Ca 가용화 능력을 검정한 결과 OPN이 CPP와 poly-glutamic acid 보다 더 우수한 것으로 나타났다.

Polymerase chain reaction을 이용한 독소생산성 Pasteurella multocida의 검출 (PCR technique for detection of toxigenic Pasteurella multocida in mixed bacterial cultures from pigs)

  • 지영철;이동석;한정희;한경수;한태욱
    • 대한수의학회지
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    • 제40권1호
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    • pp.56-62
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    • 2000
  • Pasteurella multocida is kind of commensal bacteria in the upper respiratory tract of pigs. It is classified toxigenic and nontoxigenic strains based on the production of dermonecrotic toxin. Toxigenic strain is most associated with atrophic rhinitis which brings great economical loss in swine industry. However, toxigenic and nontoxigenic strains do not differ by diagnostic biochemical reaction or morphology. One of recently developed techniques, PCR detects the toxigenic P multocida. Amplification of an 846-nucleotide fragment of toxA gene was developed. The fragment amplified by PCR was detected in P multocida type D not type A. The PCR amplification was as sensitive as it could detect 1 pg of P multocida DNA. We compared the result of the PCR with the enzyme linked immunosorbent assay (ELISA) in a test for 40 swine nasal swabs. All of these isolates were toxin negative based on the ELISA while 2 isolates were detected in the PCR technique. in addition to accuracy, as required for rapid detection from contaminated nasal swabs, toxigenic P multocida was recovered efficiently from contaminated culture without inhibition of the PCR. The results show that the PCR detection of toxigenic P multocida directly form nasal swabs are feasible.

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국내 사육 닭에서 분리된 Ornithobacterium rhinotracheale (OR)균의 약제 감수성 및 항체보유율에 대한 연구 (A Study on Antibacterial Activity and Seroprevalence of Ornithobacterium rhinotracheale Isolated from the Domestic Chickens)

  • 전우진;권용국;윤여성;김재홍
    • 미생물학회지
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    • 제39권3호
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    • pp.161-165
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    • 2003
  • Ornithobacterium rhinotracheale(OR)은 최근에 알려진 그람음성 간균으로서 가금류에서 호흡기감염을 일으킨다. OR균의 항생제 감수성과 OR에 대한 국내발생현황을 조사하기 위하여 11개 OR분리주의 8중 항생제에 대한 최소발육억제농도(MIC)와 국내 달의 OR 항체보유율을 조사하였다. 모든 분리주는 ampicillin, tetracycline 및 doxycycline에 대하여 높은 감수성을 나타내었다. MIC는 각각 0.38~2 ${\mu}g$ /ml, 0.094~3 ${\mu}g$/ml, 0.094~3 ${\mu}g$/ml으로 ciprofloxacin, norfloxacin, enrofloxacin및 ofloxacin의 MIC는 대부분 3 ${\mu}g$/ml~48 ${\mu}g$/ml로 나타났고 gentamicin에 대하여는 모든 분리주가 저항성을 나타내었다. 시판되는 OR ELISA검사 kit를 사용하여, 국내에서 사육하는 육계 및 육용종계와 산란계 188계군에 대한 항체보유율을 검사한 결과, 육계 5계군(4%),육용종계 17계군(50%),산란계 16계군(55.2%)이 OR항체 양성계군으로 나타났다. 이 결과로 보아 OR은 국내 양계장내 광범위하게 만연되어 있을 것으로 보인다.

토끼 항 ${\beta}-Lactoglobulin$ 항혈청에 대한 유청단백질 가수분해물의 항원성 (Antigenicity of Whey Protein Hydrolysates against Rabbit Anti ${\beta}-Lactoglobulin$ Antiserum)

  • 이수원;하월규;전석락;김정완;손동화;이재영
    • 한국식품과학회지
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    • 제26권5호
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    • pp.532-538
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    • 1994
  • Chymotrypsin, trypsin, pancreatin, 그리고 Aspergillus oryzae 유래 단백질분해효소의 in vitro 처리에 의하여 유청단백질(WPI)의 가수분해물(WPH)중 ${\beta}-LG$유래의 항원성변화를 조사하기 위하여 토끼 항${\beta}-LG$항혈청을 이용한 competitive inhibition ELISA(cELISA)와 heterologous PCA를 실시하였다. cELISA에 의하여 WPH의 monovalent항원성을 분석한 결과, 전체적으로 ${\beta}-LG$유래의 monovalent항원성은 효소처리에 의하여 $10^{-1.7}{\sim}10^{-4.1}$배 또는 그 이하로 저하되었으며, 특히 pepsin전처리후 Asp. oryzae유래의 효소로 가수분해한 경우(OUP)의 항원성은 거의 상실되었다. Guinea pig를 이용한 PCA test에 의하여 ${\beta}-LG$유래의 polyvalent항원성을 분석한 결과, WPH의 항원성은 $1/2{\sim}1/128$ 또는 그 이하로 저하되었다. 특히, WPH중에서 열변성이나 pepsin의 전처리없이 Asp. oryzae유래의 효소로 가수분해한 경우(OUN), 가수분해도(DH)가 그다지 높지 않고 monovalent항원성도 여전히 잔존하였음에도 불구하고($10^{-3.2}$배로 저하) 알레르기의 발증과 밀접한 관련이 있는 polyvalent항원성은 거의 상실되었다. 이는 OUN의 분해효율이 아주 높지는 않으나 ${\beta}-LG$상의 항원결정기가 효과적으로 파괴되어, polyvalent항원성이 제거되었기 때문으로 추측된다. 이 결과는, Asp. oryzae유래의 효소를 WPI에 처리하면 우유 알레르기의 주요 원인물질인 ${\beta}-LG$의 polyvalent항원성이 제거됨으로써 저알레르기성 infant formula용 WPH가 제조될 수 있음을 시사하고 있다.

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디엠프리(녹차추출물)에 의한 나균 감염 중간엽줄기세포의 IL-6 생산 억제 (DMfree®(Green Tea Extract) Inhibits IL-6 of Mycobacterium leprae Infected Mesenchymal Stem Cells)

  • 박란숙
    • 한국식품영양학회지
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    • 제28권4호
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    • pp.695-701
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    • 2015
  • Previous reports revealed that DMfree (green tea extract) inhibited expression of the IL-6 gene in Mycobacterium lepraeinfected MSCs (mesenchymal stem cells). This study aimed to measure IL-6, $IL-1{\beta}$, $TNF-{\alpha}$ and PGE2 production in M. leprae-infected MSCs using ELISA. To confirm the effect of DMfree on IL-6 and signal transduction, a western blotting test was performed. DMfree inhibited the expression of IL-6 in the MSCs and the heterodimer of STAT3, which also affects the expression of multiple genes. Though DMfree pre-treatment of control MSCs produced a baseline level of IL-6, it significantly inhibited the production of IL-6 in M. leprae-infected MSCs. There was no significant difference in IL-6 production between 1 and 7 day treatment groups. M. leprae-infected MSCs produced more $IL-1{\beta}$, $TNF-{\alpha}$ and PGE2, but DMfree could not inhibit their production at a physiological concentration. This is different from other reports that used higher concentration of EGCG treatment, resulting in significant inhibition of the cytokines. The inhibition appears to be related to the concentration of EGCG. These results indicate that DMfree can alleviate inflammation involving IL-6.

미생물효소에 의한 우유 casein의 항원성 저감화 (Reduction of Antigenicity of Bovine Casein by Microbial Enzymes)

  • 채현석;안종남;정석근;함준상;인영민;김동운
    • Journal of Dairy Science and Biotechnology
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    • 제21권2호
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    • pp.97-104
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    • 2003
  • 본 연구에서는 우유 앨러지의 원인물질중의 하나인 casein단백질을 생체가 알르겐 물질로 인식할 수 없는 수준의 분자로까지 분해하여 항원성을 저감시키고 또한 풍미가 좋은casein분해물을 제조하기 위하여 분해특성이 다른 효소를 사용하여 만들어진 분해물의 특성과 항원성을 조사하였다. 박테리아 효소인 Neutrase, Alcalase, Protamex, Pescalase을 처리한 casein가수분해물의 평균분자량은 1,100~2,300 dalton이었고 곰팡이 효소인 MP, Flavourzyme, LP, Promod경우에는 거의 770 ~ 1,140 dalton범위였다. 항원성 저감정도를 측정하기 위하여 토끼항casein항혈청을 이용하여 ELISA억제시험을 실시한 결과 항체결합을 50%저해하는 미분해casein단백질의 농도는 $10^{3.6}$ng/ml였다. 박테리아 효소인 Neutrase, Alcalase, Protamex, Pescalase에 의한 Casein가수분해물의 값은 각각 $10^{4.8},\;10^{5.5},\;10^{5.9},\;10^{6.1}\;ng/ml$이었고, 곰팡이 효소인 MP, Flavourzyme, LP, Promod경우에는 각각 $10^{6.3},\;10^{6.3},\;10^{6.5},\;10^{6.8}\;ng/ml$로 나타났다. 박테리아와 곰팡이효소의 조합인 Fla+prota, Pro+prota처리구에서는 $10^{7.4},\;10^{7.3}ng/ml$로 나타났다. 따라서 미분해 casein의 항원성을 1로 하면, casein분해물의 항원성은 최고 1/8,000 이하로 저하되었다. 수신피부아나피랙시스반응(PCA)을 이용한 항원성시험에서 casein으로 피하 감작할 경우 푸른색 반점이 나타났으나 효소처리구에서는 이러한 반점이 나타나지 않아 충분히 항원성이 저감화 되었음을 확인하였다.

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Serological responses and protection levels in chickens administered with Newcastle disease vaccines

  • Geumji Seung;Jiye Kim;Hyobi Kim;Ji-Yeon Kim;Yang-Ho Jang;Yeon-Hee Kim;Moon Her;Seong-Joon Yi;Keun-Woo Lee;Il Jang;Young Ju Lee
    • 대한수의학회지
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    • 제62권4호
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    • pp.29.1-29.7
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    • 2022
  • Vaccination against Newcastle disease (ND) is the most effective means of controlling the disease, and these vaccines are commercialized only after their safety and effectiveness have been verified through tests that comply with Korean Standards of National Lot Release for Veterinary Biologics. This study investigated whether a relatively convenient and safe serological test can be used in place of the challenge test using highly virulent ND virus. Hemagglutination inhibition (HI) assay and enzyme-linked immunosorbent assay (ELISA) were considered positive of log2 2 or more and cutoff value of 200 or more, respectively, in both live and inactivated vaccines. However, when the antibody levels of the live and inactivated vaccines induced using the Ulster 2C, KBNP-C4152R2L, and K148/08 strains were compared, the antibody titers for inactivated vaccines were significantly higher than those for live vaccines in both the HI assay and ELISA. A strong positive correlation was observed between HI and ELISA antibody titers. The live vaccines corresponded to a survival rates of ≥ 80% and the inactivated vaccines corresponded to 100% survival rates. This study confirmed that standard efficacy tests can serve as serological tests, and can replace the challenge test and that the vaccine approval process can be improved.

산란계 밀집지역의 산란저하성 질병에 관한 연구 (Studies of the egg drop laying diseases from the mass zone layer)

  • 이정원;엄성심;박인규;배정준;정동석;송희종
    • 한국동물위생학회지
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    • 제28권2호
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    • pp.121-146
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    • 2005
  • Newcastle disease (ND), infectious bronchitis (IB), low pathogenic avian Influenza (LPAI) and fowl typhoid (FT) have been known as egg drop laying diseases because of the serious layer damage from mass zone layer. In this study, such egg drop laying diseases were investigated. To access this study, we peformed to evaluate antibody titers in serum and isolated bacteria and virus from organs and feces on May, July and September in 2003. The distribution of ND from January to May, IB and LPAI from October to February of the next year, and FT from March to September were inspected by the question survey in 21 farms. ND revealed to be positive rates of 490 to 474 $(96.7\%)$ in May, 510 to 506 $(99.2\%)$ in July and 510 to 510 $(100\%)$ in September with hemagglutination inhibition (HI) test. The mean antibody titers were 10.2, 9.9 and 10.2, respectively. With regard to IB, 484 out of 490 samples $(98.7\%)$ in May, 508 of 510 $(99.6\%)$ in July and 509 of 510 $(99.8\%)$ in September showed positive results and the mean antibody titers were gradually increased with 8.2, 9.0 and 9.4, respectively. According to HI test of LPAI, the positive results were shown in 442 of 480 $(92.1\%)$, 394 of 494 $(79.8\%)$ and 402 of 483 $(83.2\%)$ in May, July and September, respectively The mean antibody titers were decreased with 4.6, 4.3 and 4.0. The distribution of LPAI also elicited the positive rates of 480 to 475 $(99.0\%)$ in May, 494 to 485$(98.2\%)$ in July, 483 to 472 $(97.7\%)$ in September as determined by ELISA and the mean S/P ratio were 2.319, 2.557 and 2.380, respectively. Compared ELISA results with HI test of LPAI the positive results were 480 to 422 $(92.1\%),\;475(99.0\%),\;494\;to\;394 (79.8\%),\;485 (98.2\%)\;and\;483\;to\;402(83.2\%),\;472(97.7\%)$. Therefore, the positive rate determined by ELISA was higher than that of HI test with 6.9, 18.4 and $14.5\%$, respectively. When performed RT-PCR for ND using organ and feces samples, the pathotypes were detected $5(15.6\%)\;in\;May,\; 2(5.3\%) in\;July,\;2(7.1\%)$ in September but there is no samples showing positive band for LPAI. In attempt to isolate Salmonella gallinarum, bacteria were obtained from 4 cases (12.5%) in May, 9 (23.6%) in July, 5 (17.8%) in September. Thus the highest rate for isolation revealed to be shown in July When evaluated the antimicrobial susceptibility to 18 isolated strains of 5. gallinarum, bacteria were sensitive to trimethoprim/sulfamethox$(61.1\%),\;kanamycin\;(55.5\%),\;ampicillin\;(55.5\%)$ and amoxacillin/clavulanic acid $(55.5\%)$, cephalothin $(50.0\%)$, but resistant to penicillin $(88.9\%)$, streptomycin $(88.9\%)$, erythromycin $(83_4\%)$ and tetracycline $(61.1\%)$. According to HI test of ND and LPAI using captured 164 wild Korean tree sparrows (Passer nontanus), the positive rates were $47.6\%\;and\;57.3\%$, and the mean HI titers were 5.32 and 4.02, respectively. 71 $(43.2\%)\;and\;58(35.3\%)$ in captured sparrows also showed more than 4 titers for HI test to ND and LPAI, respectively However, the attempt for isolation of viruses failed in all samples.