• 제목/요약/키워드: ELISA method

검색결과 593건 처리시간 0.03초

Comparison of Gold Biosensor Combined with Light Microscope Imaging System with ELISA for Detecting Salmonella in Chicken after Exposure to Simulated Chilling Condition

  • Mi-Kyung Park
    • Journal of Microbiology and Biotechnology
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    • 제33권2호
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    • pp.228-234
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    • 2023
  • In this study, the performance of a gold biosensor combined with light microscope imaging system (GB-LMIS) was comparatively evaluated against enzyme-linked immunosorbent assay (ELISA) for detecting Salmonella under simulated chilling condition. The optimum concentration of antiSalmonella polyclonal antibodies (pAbs) was determined to be 12.5 and 100 ㎍/ml for ELISA and GBLMIS, respectively. GB-LMIS exhibited a sufficient and competitive specificity toward three tested Salmonella among only. To mimic a real-world situation, chicken was inoculated with Salmonella cocktail and stored under chilling condition for 48 h. The overall growth of Salmonella under chilling condition was significantly lower than that under non-exposure to the chilling condition (p < 0.05). No significant differences in bacterial growth were observed between brain heart infusion and brilliant green broth during the enrichment period (p > 0.05). Finally, both GB-LMIS and ELISA were employed to detect Salmonella at every 2-h interval. GB-LMIS detected Salmonella with a competitive specificity by the direct observation of bacteria on the sensor using a charge-coupled device camera within a detection time of ~2.5 h. GB-LMIS is a feasible, novel, and rapid method for detecting Salmonella in poultry facilities.

살균제 Iprovalicarb 잔류물의 신속한 검출을 위한 효소면역분석법 (Enzyme Immunoassay for Rapid Detection of the Fungicide Iprovalicarb Residues)

  • 조한근;경기성;이은영
    • Journal of Biosystems Engineering
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    • 제31권6호
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    • pp.535-540
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    • 2006
  • For a biosensor development, an enzyme-linked immunosorbent assay (ELISA) of the fungicide iprovalicarb was developed by minimizing the processing time. The time for whole incubation process was reduced from 135 minutes to 15 minutes. The concentration of antibody was varied to improve sensitivity. The total processing time was reduced from 2.5 hours to 20 minutes, the final sensitivity ($IC_{50}$ value) of 7.93 ng/mL and the lowest detection limit of 0.045 ng/mL were obtained. This ELISA was applied to potatoes and onions, and the recoveries were in the range of 98.85 $\sim$ 101.20% and 87.97 $\sim$ 102.70%, respectively. Accordingly, this method can be used as basis for a biosensor for rapid monitoring of iprovalicarb residues in crops.

Resorcin Blue 염색 기법에 의한 감자 잎말림병의 형광 현미경적 진단 (Diagnosis of Potato Leafroll disease by Fluorescence Microscopic Detection of Callose Stained with Resorcin Blue)

  • 이철호;나용준
    • 한국식물병리학회지
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    • 제11권2호
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    • pp.101-106
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    • 1995
  • Deep blue fluorescence of resorcin blue-stained callose was observed only in the potato leafroll virus (PLRV)-infected potato plants, but not in other potato viruses investigated. The plant sections stained with aniline blue showed non-specific fluorescence regardless of PLRV infection, which means that aniline blue is not suitable for the staining of callose induces by PLRV infection. The fluorescence of resorcin blue-stained callose was more easily detectable than autofluorescence by a direct fluorescence detection method because of its deep blue color. The lateral branch of lower leaves was turned out to be the best material for fluorescence observation of all plant parts tested. In comparison of diagnostic efficacy of this technique to enzyme-linked immunosorbent assay (ELISA), PLRV infected potato plants showed corresponding increment of the fluorescence of resorcin blue stained callose as absorption values in ELISA increased. In the future, the criteria measuring the fluorescence objectively are thought to be determined for the practical application to the diagnosis of potato leafroll disease.

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Antibody derived from insect glycosaminoglycan

  • Ahn, Mi Young;Hwang, Jae Sam;Yoon, Hyung Joo;Yun, Eun Young
    • International Journal of Industrial Entomology and Biomaterials
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    • 제29권2호
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    • pp.214-219
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    • 2014
  • We prepared antibodies from insect glycosaminoglycans (GAGs) and assayed the titer. Nine polyclonal antibodies against insect GAGs were raised for development of an ELISA in biological fluids (mice serum). The 3th booster collection of antiserum of BALB/c mice as a primarily antibody was assayed for titer determination by ELISA method. In sandwich ELISA of GAGs derived from Isaria sinclairii or other insects, antiserum from insect GAGs gave satisfactory results for so potent antibody(100: 1~1000:1) raising (manufacturing) agent in range of 10 ng/ml.

Development of a Disperse Dye Immunoassay Technique for Detection of Antibodies against Neospora caninum in Cattle

  • Selahi, Fatemeh;Namavari, Mehdi;Hosseini, Mohammad Hossein;Mansourian, Maryam;Tahamtan, Yahya
    • Parasites, Hosts and Diseases
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    • 제51권1호
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    • pp.129-132
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    • 2013
  • In this study a disperse dye immunoassay method was standardized and evaluated for detection of antibodies against Neospora caninum in cattle. Sera from 150 cattle with a recent history of abortion were collected and tested by commercial ELISA kit and a standardized in-house dye immunoassay system. The positivity rate for the sera used in this study was 34.6% for the disperse dye immunoassay (DDIA) compared to 32% obtained by ELISA kit. This study showed no significant difference between DDIA and ELISA. The results indicated that the DDIA provide an economic, simple, rapid and robust test for detection of N. caninum infection in cattle.

Serum Antigen and Antibody Detection in Echinococcosis: Application in Serodiagnosis of Human Hydatidosis

  • Sadjjadi, Seyed Mahmoud;Sedaghat, Farzaneh;Hosseini, Seyed Vahid;Sarkari, Bahador
    • Parasites, Hosts and Diseases
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    • 제47권2호
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    • pp.153-157
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    • 2009
  • Diagnosis of hydatidosis is based on immunodiagnostic methods along with radiological and ultrasound examinations. The objectives of the present study were to develop a specific and simple antigen-based ELISA method for diagnosis of hydatidosis and compare it with antibody detection method. The subjects in this study included 89 patients in the following groups: surgically confirmed hydatidosis patients (35 cases), control with other parasitic diseases (29 cases), and healthy controls (25 cases). Hyperimmune serum was raised against hydatid cyst fluid in rabbits. Anti-hydatid cyst IgG was purified by affinity chromatography using protein A column and labeled with horseradish peroxidase. Collected sera were assessed for hydatid cyst antigens and antibody by ELISA. Circulating hydatid antigen was found in 9 out of 35 patients with surgically confirmed hydatidosis. A sensitivity of 25.7% and a specificity of 98.0% were calculated for the antigen detection assay. Antibody detection by indirect ELISA, using antigen B, showed that 94.2% of patients (33 cases) have anti-hydatid cyst antibodies in their serum while cross reaction was noted in a few of non-hydatidosis patients. A sensitivity of 94.2% and specificity of 81.6% were found for the antibody detection assay. Findings of this study indicated that antibody detection assay is a sensitive approach for diagnosis of hydatid cyst while antigen detection assay might be a useful approach for assessment of the efficacy of treatment especially after removal of the cyst.

Haptoglobin Concentration in the Cord Blood of Uninfected Korean Newborns

  • Choi, Seong Jin;Lee, Byoungkook;Ahn, Kwangjin;Uh, Young
    • Perinatology
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    • 제29권4호
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    • pp.165-169
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    • 2018
  • Objective: Highly sensitive haptoglobin measurement should be used in neonates because the haptoglobin concentration in neonates is lower than that of adults. The aim of this study was to establish the reference values of haptoglobin levels in the cord blood of uninfected neonates. Methods: The cord blood of 29 preterm and 51 term babies was collected, and data from the mother and the newborn were recorded. The haptoglobin concentrations of 80 cord blood samples were simultaneously measured by enzyme-linked immunosorbent assay (ELISA; Assaypro, St Charles, MO, USA) and immunoturbidimetry assay (Roche Diagnostics, Basel, Switzerland). C-reactive protein (CRP) was also measured by immunoturbidimetry assay (Roche Diagnostics, Switzerland). Results: Mean values of CRP and ELISA haptoglobin were not significantly different between preterm and term babies. The 2.5 percentile and 97.5 percentile values of ELISA haptoglobin concentration were as follows: 80 neonates, 0.01 mg/dL and 0.59 mg/dL; 29 preterm babies, 0.08 mg/dL and 0.18 mg/dL; and 51 term babies, 0.07 mg/dL and 0.23 mg/dL. There were no differences in ELISA haptoglobin concentration according to maternal underlying diseases, delivery method, usage of antibiotics or steroids before delivery, gestational age, gender of baby, or twin gestation. Conclusion: A highly sensitive haptoglobin method should be used to determine the haptoglobin concentration in Korean newborns because the reference values of cord blood haptoglobin concentration in Korean newborns are less than the lower detection limit for commonly used immunoturbidimetric haptoglobin measurement methods.

Method validation for quantitative analyzing aflatoxin productivity in Aspergillus sp. isolated from soybean paste

  • SeongEui Yoo;WooSoo Jeong;Soo-Hwan Yeo;So-Young Kim
    • 한국식품저장유통학회지
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    • 제30권1호
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    • pp.28-41
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    • 2023
  • Non-aflatoxigenic Aspergillus oryzae and aflatoxigenic A. flavus cannot be clearly identified by partial sequencing of the internal transcribed spacer (ITS) and 18S ribosomal ribonucleic acid (18S rRNA) regions. This study aimed to compare the accuracy among three aflatoxin detection methods using ultra-performance liquid chromatography (UPLC), high-performance liquid chromatography (HPLC), and an enzyme-linked immunosorbent assay (ELISA) kit and to select the non-aflatoxigenic Aspergillus sp. isolated from soybean paste. All analytical methods were suitable according to the international standards of Codex Alimentarius FAO-WHO (CODEX) or the Ministry of Food and Drug Safety (MFDS). UPLC exhibited the best of limit of detection (LOD) and limit of quantification (LOQ). Based on UPLC, HPLC, and the ELISA kit assay, the P5 and P7 strains isolated from soybean paste had 1,663.49, 1,468.12, and >20 ㎍/kg and 1,470.08, 1,056.73, and >20 ㎍/kg, respectively, detected and re-identified as A. flavus. In contrast, the P3 and P4 strains (A. oryzae), which were detected below the MFDS standards in all assays, were confirmed as non-aflatoxigenic fungi. Among the methods evaluated for quantitative analysis of aflatoxin, UPLC and HPLC are superior in terms of accuracy, and the ELISA kit rapidly detects low concentrations of aflatoxin. Furthermore, this study demonstrates that any Aspergillus sp. isolated for use as a fermentation starter should be analyzed for potential aflatoxin production using UPLC and HPLC for accurate quantitative analysis or ELISA for the rapid detection of low-level concentrations of aflatoxin.

영남지방 농상물에 대한 위생학적 연구(제2보) ELISA 법에 의한 Aflatoxin B1 검색 (Hygienic Studies on the Agricultural Products in Youngnam Districts (Part II) Determination of Aflatoxin B1 by ELISA Method)

  • 정덕화;하기수;정혜경;김동술
    • 한국식품위생안전성학회지
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    • 제4권3호
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    • pp.171-176
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    • 1989
  • 영남지방 곡류에 오염되어 있는 Aflatoxin B1을 측정하기 위해 식속하고 정확한 Enzyme-Linked Immunosorbent Assay(ELISA) 법을 응용하였다. 생산된 Antibody 는 aflatoxin B2와 강한 corss reaction을 보였고, 정량적으로 표준곡선을 보인 aflatoxin B1 범위는 1-100ppb이었으며, methanol-PBS-dimethylformamide로 spike시킨 쌀 추출물이 ELISA 반응에 약간의 간섭반응을 보였다. ELISA법에 의한 쌀 65종, 보리 116종의 aflatoxin 분석결과는 다음과 같다. 즉, 쌀시료 65개 중 3개의 시료가 positive 반응을 보였으며, 그중 R-18이 $7.5\;\mu\textrm{g}/kg$, R-30이 $3.6\;\mu\textrm{g}/kg$, R-59가 $3.3\;\mu\textrm{g}/kg$ 순으로 aflatoxin B1 함량을 나타내었고 시료의 4.6%가 aflatoxin B1에 오염되었음을 보였다. 보리 116종 중 겉보리에서 4개의 시료가 positive 반응을 나타내었으며, 그 중 겉보리 UB-37이 $9.6\;\mu\textrm{g}/kg$의 함량으로 가장 높은 함량을 나타내었고, 그의 UB-35, UB-54순이었으며 전체 시료의 3.4%가 aflatoxin B1에 오염되었음을 보였다.

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