• Title/Summary/Keyword: ELISA method

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A study on the massive cultivation of cyanobacteria and strip analysis of cyanobacterial toxin (남조류의 대량배양 및 남조류 독소의 스트립분석법 연구)

  • Pyo, Dongjin;Yim, Miyeon;Kim, Eujin
    • Analytical Science and Technology
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    • v.25 no.6
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    • pp.388-394
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    • 2012
  • Cyanobacterial toxins, microcystins which exist in Korean lakes show strong toxicity to fish, cattles and human. In this study, we tried to analyze cyanobacterial toxin, microcystin in the Microcystis cultivation solution using test strip, although the most common analytical methods for the detection of microcystin are HPLC and ELISA. This new anlytical method used the advantages of high specifisity and rapidness of test strip, high sensitivity of fluorescence reader. Therefore, we could analyze the trace amount of microcystin existed in various water samples without using the microcystin standards.

Dietary Exposure of Aflatoxin$B_1$ and Cancer Risk Assessment (아플라톡식 $B_1$ 노출에 의한 발암 위해성 평가)

  • 이병무;최문정;변수현;김형식
    • Journal of Food Hygiene and Safety
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    • v.10 no.2
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    • pp.81-87
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    • 1995
  • Daily exposure of aflatoxin B1 (AFB1) was estimated in foods (rice, barley, soybean, peanut, soysauce, soybean paste) by ELISA (enzyme linked immunosorbent assay) using polyclonal antibody R101. Before ELISA, a simple extraction method was applied for the quantitation of AFB1 in foods using chloroform which showed high recovery (70$\pm$12%). AFB1 levels in foods were 0.32 ng/ml (rice), 0.24ng/ml (barley), 0.22 ng/ml (peanut), 0.30~0.78 ng/ml (soysauce), and 0.2 ng/ml (soybean paste). Based on food consumption, we estimated that Koreans were exposed to AFB1 at the level of 1.86$\pm$0.46 ng/kg/day and liver cancer incidence attributed to AFB1 exposure (assuming that AFB1 as a single hepatocarcinogenic agent) might be calculated to be 13.1 per 100, 000 population. Our data demonstrate that AFB1 levels in foods were below the regulation of 10 ppb in foods and might not be the major risk factor for the high incidence of lover cancer in Korea.

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Prevalence and Characterization of Enterohemorrhagic Escherichia coli (EHEC) Isolated from Ground Beefs Distributed in Gyeong-In Region (경인지역에 유통되는 분쇄육 중 장출혈성대장균의 분포 조사 및 특성 연구)

  • Kim, Eun-Jeong;Park, Yong-Chjun;Cho, Joon-Il;Lee, Jong-Ok;Kim, Hee-Yun
    • Korean Journal of Food Science and Technology
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    • v.38 no.6
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    • pp.773-778
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    • 2006
  • The objective of this study was to evaluate three verocytotoxin-producing Escherichia coli (VTEC) detection kits to detect the presence of VT genes: Doupath Verocytotoxin (GLISA) developed by MERCK, ProsPect Shiga Toxin E. coil (STEC) Microplate Assay (ELISA) developed by Remel, and a polymerase chain reaction method. Our laboratory verified artificially inoculated samples. All three methods could detect very low numbers of VTEC, but VT-PCR had the best sensitivity for VTEC detection. From April through September 2005, 257 ground-beefs from supermakets and traditional markets were examined for the presence of VTEC by polymerase chain reaction immediately after purchase and total viable counts (TVC) were determined. VTEC was isolated from 30 of 257 ground-beefs. A variety of serogroups was found, including 10 stains belonging to the virulence type EHEC, but major serogroups such as O157, O26 and O111 were nor found.

An Effective Method of Diagnosis of Potato Leafroll Virus by RT-PCR (RT-PCR 방법을 이용한 효과적인 감자 잎말림 바이러스의 검정)

  • Jeon, Jae-Heung;Joung, Young-Hee;Choi, Kyung-Hwa;Kim, Hyun-Soon;Oh, Hyun-Woo;Park, Se-Won;Joung, Hyouk
    • Korean Journal Plant Pathology
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    • v.12 no.3
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    • pp.358-362
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    • 1996
  • 감자 잎말림 바이러스를 검정하기 위하여 ELISA 및 전자현미경에 의해 바이러스 감염이 확인된 기내 배양중인 감장의 줄기로부터 RT-PCR 분석을 수행하였다. 분리된 총 RNA들로부터 바이러스 cDNA를 합성하고 감자 잎말림 바이러스 외피단백질의 일부인 465bp를 특이하게 증폭하도록 고안한 두 primer를 사용하여 PCR 반응을 하였다. 증폭된 465pb의 DNA 절편의 염기서열을 분석한 결과 역시 감자 잎말림 바이러스임을 확인하였다. 바이러스 검정에 있어서 EL-ISA 방법과 RT-PCR 방법간의 민감도를 조사한 결과 RT-PCR 방법간의 민감도를 조사한 결과 RT-PCR 방법이 ELISA 방법보다 감자 잎말림 바이러스검정에 있어서보다 정확한 방법인 것으로 사료된다.

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Genetic Diversity in the Coat Protein Genes of Prune dwarf virus Isolates from Sweet Cherry Growing in Turkey

  • Ozturk, Yusuf;Cevik, Bayram
    • The Plant Pathology Journal
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    • v.31 no.1
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    • pp.41-49
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    • 2015
  • Sweet cherry is an important fruit crop with increasing economical value in Turkey and the world. A number of viruses cause diseases and economical losses in sweet cherry. Prune dwarf virus (PDV), is one of the most common viruses of stone fruits including sweet cherry in the world. In this study, PDV was detected from 316 of 521 sweet cherry samples collected from 142 orchards in 10 districts of Isparta province of Turkey by double antibody sandwich-enzyme linked immunosorbent assay (DAS-ELISA). The presence of PDV in ELISA positive samples was confirmed in 37 isolates by reverse transcription- polymerase chain reaction (RT-PCR) method. A genomic region of 862 bp containing the coat protein (CP) gene of PDV was re-amplified from 21 selected isolates by RT-PCR. Amplified DNA fragments of these isolates were purified and sequenced for molecular characterization and determining genetic diversity of PDV. Sequence comparisons showed 84-99% to 81-100% sequence identity at nucleotide and amino acid level, respectively, of the CP genes of PDV isolates from Isparta and other parts of the world. Phylogenetic analyses of the CP genes of PDV isolates from different geographical origins and diverse hosts revealed that PDV isolates formed different phylogenetic groups. While isolates were not grouped solely based on their geographical origins or hosts, some association between phylogenetic groups and geographical origins or hosts were observed.

Development of serodiagnostic surface plasmon resonance imaging assay for the detection of antibodies to porcine circovirus type 2

  • Park, Chul;Kim, Bum-Seok;Kim, Yong-Hwan;Cho, Ho-Seong
    • Korean Journal of Veterinary Service
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    • v.34 no.1
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    • pp.1-4
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    • 2011
  • A surface plasmon resonance imaging (SPRI) assay was developed for measuring porcine circovirus type 2 (PCV2) antibody using a recombinant capsid protein as an antigen. The diagnostic potential of SPRI for detecting antibodies to the PCV2 capsid protein was compared with that of a conventional enzyme-linked immunosorbent assay (ELISA) using 70 pig serum samples taken from 6 pig farms. There was a strong positive correlation between the SPRI and ELISA (n = 70, r = 0.911, P<0.01). Therefore, this recombinant capsid protein can be used as an antigen for serological studies, and the SPRI, a label-free and high-throughput method, is expected to be a valuable tool in the serodiagnosis of PCV2 infection.

Effects of Draconis Resina on the Collagenase Activities and the Procollagen Synthesis in Hs68 Human Fibroblasts, and Tyrosinase Activity

  • Kim, Tae Yeon;Leem, Kang-Hyun
    • The Korea Journal of Herbology
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    • v.30 no.6
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    • pp.1-6
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    • 2015
  • Objectives : Draconis Resina (DR), the resin of Daemonorops draco Bl., is used to circulate the blood and to stop bleeding. It also has been used to generate flesh including ulceration. The present study investigated the effects of DR extract on collagen metabolism in human fibroblasts and tyrosinase activity in mushroom tyrosinase.Methods : The effect of DR extract on type I procollagen production (collagen type I synthesis) and collagenase (matrix metalloproteinase-1, henceforth referred as MMP-1) activity in human normal fibroblasts cell line. Hs68 cells after ultraviolet B (UVB, 312 nm) irradiation was measured using the enzyme - linked immunosorbent assay (ELISA). The tyrosinase activity was also measured to find out the whitening effects in mushroom tyrosinase by ELISA method.Results : There was no cytotoxicity at DR extract at concentrations of 10 μg/ml, 30 μg/ml, and 100 μg/ml. DR extract significantly inhibited the increase of collagenase activity, whereas it did not show on the reduction of type I procollagen in UVB damaged Hs68 cells. DR extract did not reduce the L - DOPA oxidation. However, it significantly reduced the tyrosinase activity by DR extract at concentraions of 0.1 mg/ml, 1 mg/ml and 10 mg/ml.Conclusions : In conclusion, DR showed the anti-wrinkle and whitening effects via the inhibition of collagenase production and the tyrosinase activity. These results suggest that DR may have potential as an anti-aging ingredient in cosmetic herb markets.

Identification of the IL-1$\beta$ inhibitor in the febrile patient urine by anti-IL-1$\beta$ monoclonal antibody (Anti-IL-1$\beta$ 단일클론 항체를 이용해서 발열환자의 뇨중 IL-1$\beta$ inhibitor의 확인)

  • 남경수;배윤수;남명수;오은숙;박순희;최인성;정태화
    • YAKHAK HOEJI
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    • v.37 no.4
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    • pp.420-426
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    • 1993
  • To effectively purify of IL-1 inhibitor from human febrile urine, we have established monoclonal antibody that reacts with human recombinant interleukin l$\beta$(IL-1$\beta$). The antibody, designated ON-1, was highly specific to IL-1$\beta$ and no cross-reaction with other cytokines(IL-l$\alpha$ and IL-4) was observed. As the results of ELISA inhibition assay and Western blotting method, it was further identified that ON-1 had high binding specificity with IL-1$\beta$. IL-1 receptor binding material from febrile patient urine was effectively purified with affinity column chromatography which conjugated with ON-1. This urinary material inhibited the thymocyte proliferation in a dosedependent manner. IL-l$\beta$ induced thymocyte proliferation activity was inhibited to 67.3% at 6 $\mu\textrm{g}$ of the purified urinary material. The result may suggest that this urinary material the purified urinary material. The result may suggest that this urinary material will have antagonic effect on IL-1 action mechanism and act IL-l$\beta$ inhibitor.

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Antiserum Preparation of Recombinant Sweet Potato Latent Virus-Lotus (SPLV-Lotus) Coat Protein and Application for Virus-Infected Lotus Plant Detection

  • He, Zhen;Dong, Tingting;Chen, Wen;Wang, Tielin;Gan, Haifeng;Li, LiangJun
    • The Plant Pathology Journal
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    • v.36 no.6
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    • pp.651-657
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    • 2020
  • Lotus is one of the most important aquatic vegetables in China. Previously, we detected sweet potato latent virus from lotus (SPLV-lotus) and found that it has highly significant sequence diversity with SPLV-sweet potato isolates (SPLV-sp). Here, we developed serological methods for the detection of SPLV-lotus in Chinese lotus cultivation areas. Based on the high sensitivity of SPLV-lotus coat protein antiserum, rapid, sensitive and large-scale diagnosis methods of enzyme-linked immunosorbent assay (ELISA) and dot blot in lotus planting area were developed. The established ELISA and dot blot diagnostic methods can be used to detect SPLV-lotus from samples successfully. And our results also showed that the SPLV-lotus and sweet potato isolates appeared clearly distinction in serology. Our study provides a high-throughput, sensitive, and rapid diagnostic method based on serology that can detect SPLV on lotus, which is suggested to be included in viral disease management approach due to its good detection level.

Sodium Hypochlorite Solution As a Chemical Wounding Agent for Improving Agrobacterium-mediated Chinese Cabbage Seed Transformation (Sodium hypochlorite처리에 따른 배추종자의 Agrobacterium이용 형질전환 증대)

  • Shin Dong-Il;Park Hee-Sung
    • Journal of Life Science
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    • v.15 no.6 s.73
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    • pp.1034-1036
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    • 2005
  • Chinese cabbage (Brassica campestris ssp. napus var. pekinensis Makino) seeds/seedlings were transformed via vacuum-infiltration with recombinant Agrobacterium tumefaciens LBA4404 cells. The agroinfiltration method was determined to be unsuccessful for Chinese cabbage transformation during the analysis of hepatitis B surface antigen expression by ELISA. However, treatment of sodium hypochlorite solution, prior to agroinfiltration, to pregerminated or germinating 1 day- or 2 days-old seeds was proven effectively to enhance transformation efficiency, suggesting that chemical wounding caused by sodium hypochlorite reaction might facilitate Agrobacterium infection and, therefore, transient gene expression in Chinese cabbage sprouts.