• Title/Summary/Keyword: Enzyme Linked Immunosorbent Assay Kit

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Synthetic Peptide-Based Enzyme-Linked Immunosorbent Assay for Human $\alpha$-Fetoprotein

  • Yoon, Mi-Chung;Lee, Hyun-Hee
    • Biomedical Science Letters
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    • v.7 no.3
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    • pp.103-110
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    • 2001
  • $\alpha$-Fetoprotein(AFP) is a good marker for the detection of several diseases such as hepatocellular carcinoma, gonadal germ cell tumor, gastric tumor, and Down's syndrome. In this study, we developed ELISA, using synthetic peptides corresponding to the epitopes of AFP. Five kinds of peptides were synthesized from AFP to produce antibodies in rats that recognize AFP in human plasma as well as amniotic fluid and do not cross-react with serum albumin. All five kinds of antibodies showed good reactivities with their peptide-keyhole limpet hemocyanin conjugates. Anti-synthetic peptide 1 (R-N-E-Y-G-I-A-S-I-L, 4-13) antibody, in particular, reacted well with AEP as well as synthetic peptide 1-KLH but not with human serum albumin. The binding affinity(Kd) was 2.7$\times$10$^{-9}$M for peptide 1 and 6.8$\times$10$^{-8}$M for AEP. The range for measurement of AFP was 10~1,000 ng/ml. The within-assay and between-assay coefficients of variance(CV) were 4.83% and 10.97%, respectively. In a sample of 31 sera and 33 amniotic fluids, there was a good correlation between AFP values determined in this assay and those in a commercial kit. These results indicate that the antibodies against synthetic peptides corresponding to the epitopes of AFP are highly specific to APP and synthetic peptide-based ELISA would be useful for the measurement of human AFP.

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High seroprevalence of Ornithobacterium rhinotracheale in layer chickens in Gyeonggi province, South Korea (경기도 산란계에서 Ornithobacterium rhinotracheale의 높은 항체 양성률)

  • Jung, Kwang
    • Korean Journal of Veterinary Service
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    • v.43 no.4
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    • pp.257-259
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    • 2020
  • Ornithobacterium rhinotracheale (ORT) causes pneumonia, airsacculitis, and pleuritis in chickens and other avian species. Little is known about the seroprevalence of ORT in layer chickens in Gyeonggi province, South Korea. The purpose of this study was to determine the seroprevalence of ORT in layer chickens in Gyeonggi province, South Korea from May to September 2019. A total of 460 chickens in 28 flocks were tested for antibodies to ORT by using commercial enzyme-linked immunosorbent assay (ELISA) kit. The seroprevalence of ORT antibodies in the flocks was 100% (28/28) and the overall seroprevalence in individual chickens was 98.91% (455/460). This survey indicated the high seroprevalence of ORT in layer chickens in Gyeonggi province, South Korea. Therefore, measures should be executed to control ORT in layer chickens in this province.

Clinical Characteristics of Pertussis Epidemic in Changwon (창원에서 유행한 백일해의 임상적 특징)

  • Jang, Joon Weon;Kang, Jin Han;Choi, Jae Won;Lee, Hak Sung;Ma, Sang Hyuk
    • Pediatric Infection and Vaccine
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    • v.24 no.1
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    • pp.37-43
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    • 2017
  • Purpose: Pertussis can be prevented with a vaccine. Despite this, there have been an increasing number of cases worldwide, and also in Korea. This study aimed to investigate the epidemiology and clinical characteristics of the recent outbreak in the Changwon area. Methods: Patients who visited Changwon Fatima Hospital from July 2015 to March 2016 with respiratory symptoms, including spasmodic cough, cough induced vomiting, inspiratory 'intake' sound (whooping), and a night-time cough for >1 week were included in this study. Respiratory specimens were collected from patients and a polymerase chain reaction (PCR) and detected anti-pertussis immunoglobulin G enzyme-linked immunosorbent assay kit test were performed. Patients with underlying diseases, or those who had received a DTaP or Tdap vaccination in recent 1 year were excluded. Results: Pertussis was diagnosed in 37 of 50 patients, two patients were positive according to the PCR, and 37 patients were positive according to serologic tests. The age distribution of the patients was 1 month to 15 years. After administering antibiotics, all patients recovered without complications. Conclusions: A pertussis outbreak occurred in Changwon in 2015 and 2016. This data can provide the basis for further study on the epidemiology of pertussis in Korea.

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

  • SeongEui Yoo;WooSoo Jeong;Soo-Hwan Yeo;So-Young Kim
    • Food Science and Preservation
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    • v.30 no.1
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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.

Comparison between indirect immunofluorescent antibody (IFA) test and enzyme-linked immunosorbent assay (ELISA) for the detection of antibody to porcine reproductive and respiratory syndrome virus(PRRSV) (돼지 생식기호흡기증후군 바이러스항체 검색에 있어 간접형광항체법(IFA) 과 효소면역법(ELISA)의 진단효율 비교)

  • Park, Choi-kyu;Lyoo, Young-soo;Lee, Chang-hee;Jung, Jong-wook
    • Korean Journal of Veterinary Research
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    • v.38 no.2
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    • pp.314-318
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    • 1998
  • An establishment of effective control measures to PRRSV infection in swine industry depends on a sensitive and specific diagnosis to detect either viral antigen and/or antibodies to PRRSV. Several diagnostic methods are available to detect antibodies against PRRSV, including IPMA, IFA and ELISA tests have been successfully developed. Sensitivity of the indirect immunofluorescent assay in MA-104 cells using Korean field isolate PL96-1 was superior to that of VR-2332 and field isolate PL96-2. Sensitivity and specificity of the IFA test with PL96-1 were comparable to those of commercial ELISA test kit but ELISA test was more sensitive for the detection of declining antibodies to PRRSV in finishing pigs. In this study we concluded that IFA and ELISA test could be utilized to detect antibodies to PRRSV and the results generated from these two tests were comparable and there were no significant difference between these two tests.

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Effect of Hwang-Ryeon-Hae-Dok-Tang on the Release of IL-8 in Human Nasal Mucosal Fibroblast (黃連解毒湯의 사람 비점막 섬유아세포 IL-8 분비에 대한 효과)

  • Lee, In-su;Kim, Hee-taek;Lee, Eun-yong;Kim, E-hwa;Ryu, Ju-hyun
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.16 no.3
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    • pp.68-81
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    • 2003
  • It is proposed that Hwang-Ryeon-Hae-Dok-Tang may modulate the immune response on allergy or asthma. Human nasal mucosal fibroblasts are a rich source cytokines, inflammatory mediators, and chemokines. Chemokines are important for the recruitment of leukocytes to sites of infection, which is essential in host defense. Objectives : The objective of this study was to investigate the effect of Hwang-Ryeon-Hae-Dok-Tang(HH) on the release of the IL-8 chemokine in human nasal mucosal fibroblasts after stimulation with cytokines like interleukin-4(IL-4), tumor necrosis factor- (TNF- ), interferon- (lFN- ), and interle ukin-l (IL-I ). Methods : To detect the release of IL-8, enzyme-linked immunosorbent assay(ELISA) kit was performed. The cytotoxicity was measured by MTT assay. Results : HH significantly inhibited the secretion of IL-8 with a dose-dependant manner. The effective dosage did not have the cytotoxicity on human nasal mucosal fibroblasts Conclusions : Results of our study show that HH would play an important role in modulation of IL-8 in human nasal mucosal fibroblasts.

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Inhibition of MMP-2 and -9 by Crude Extracts and Their Solvent-partitioned Fractions from the Halophyte Atriplex gmelinii (가는갯능쟁이(Atriplex gmelinii) 추출물과 용매분획물의 MMP-2와 MMP-9 활성 저해효과)

  • Park, Min Jeong;Kim, Junse;Kong, Chang-Suk;Seo, Youngwan
    • Ocean and Polar Research
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    • v.41 no.2
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    • pp.79-88
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    • 2019
  • In this study, the inhibitory effect of Atriplex gmelinii C. A. Mey. against the activity of MMP-2 and MMP-9 secreted from phorbol-12-myristate-13-acetate (PMA)-stimulated HT-1080 cells was evaluated by gelatin zymography and enzyme-linked immunosorbent assay (ELISA), reverse transcription polymerase-chain reaction (RT-PCR), and Western blot assay. Specimens of the halophyte A. gmelinii were extracted twice for 24 hr with methylene chloride ($CH_2Cl_2$), and then twice with methanol (MeOH), in turn. Each extract significantly inhibited the enzymatic activities in gelatin zymography and MMP ELISA kit, and expression of MMP-2 and 9 in mRNA and protein levels. Two crude extracts were combined and then the combined crude extracts were fractionated into n-hexane, 85% aqueous methanol (85% aq.MeOH), n-butanol (n-BuOH), and water ($H_2O$) fractions, according to solvent polarity. Among solvent-partitioned fractions, the 85% aq.MeOH fraction showed the strongest inhibitory effect against MMP-2 and -9 in gelatin zymography and MMP ELISA kit. In RT-PCR, all solvent-partitioned fractions significantly suppressed mRNA expression of MMP-2 and -9. On the other hand, in Western blot assay, all solvent-partitioned fractions except $H_2O$ significantly reduced expression levels of protein. HT 1080 cell migration was most significantly inhibited by the n-BuOH fraction followed by the 85% aq.MeOH and $H_2O$ fractions. These results suggest that A. gmelinii could be used as a potential source to inhibit tumor cell metastasis.

Improvement of anti-oxidant and anti-inflammatory activities of aleurone layer extracts of black rice (Oryza sativa L.) by enzyme treatment (효소 처리에 의한 흑미 호분 추출물의 산화방지와 항염증 활성 증진)

  • Lee, Mi Kyoung;Ryu, Soo In;Lee, Min Ho
    • Korean Journal of Food Science and Technology
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    • v.50 no.5
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    • pp.528-534
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    • 2018
  • The current study investigated the effects of enzyme treatment on black rice (Oryza sativa L.) aleurone layer extracts. Different enzymes (lipase, lecitase ultra and lipopan 50BG) were used to test anti-oxidant and anti-inflammatory activities in vitro. The antioxidant activities of enzyme treated or non-enzyme treated extracts of black rice bran were evaluated via 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) radical scavenging activity. Lipase treated extracts of black rice bran showed higher antioxidant activity compared to that of non-enzyme treated extracts. Anti-inflammatory activities of enzyme treated black rice bran extracts on nitrite production and tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$) secretion, were tested using a nitric oxide (NO) colorimetric assay kit and an enzyme-linked immunosorbent assay (ELISA) kit. The ethanolic extract of enzyme treated black rice bran decreased the levels of nitric oxide production and pro-inflammatory cytokines, such as tumor necrosis $factor-{\alpha}$, in a lipopolysaccharidestimulated RAW 264.7 cell culture. These findings indicate that enhanced anti-oxidant and anti-inflammation activities of the ethanolic extracts of enzyme treated black rice (Oryza sativa L.) aleurone layers, may be attributed to molecular conversion of ingredients in enzyme catalyzed reactions.

Investigation of bovine leukemia virus infection in dairy farms of northern Gyeonggi province, Korea (경기도 북부지역 젖소 사육농장의 bovine leukemia virus 감염 실태 조사)

  • Jung, Kwang;Shim, Hang-Sub;Baek, Jin-Joo
    • Korean Journal of Veterinary Service
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    • v.35 no.4
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    • pp.333-337
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    • 2012
  • This study was carried out to investigate the prevalence of bovine leukemia virus (BLV) infection and to compare the results of enzyme-linked immunosorbent assay (ELISA) and nested polymerase chain reaction (nPCR) in dairy farms in northern Gyeonggi province from August through December 2011. A total of 625 dairy cattle from 14 dairy farms were tested for antibodies against BLV using commercially available ELISA test kit. The overall seroprevalence of BLV infection was 76.3%. The seroprevalence of diary cattle according to age was the highest at 61~72 months (88.0%, P<0.001). Two hundred fifty one dairy cattle from 7 diary farms were tested ELISA and nPCR. The kappa value of BLV between ELISA and nPCR was 0.765. The results indicate that BLV infection spread widely in dairy farms and the nPCR is rapid method for the early detection of BLV infection.

The Characteristics, Detection and Control of Bacteriophage in Fermented Dairy Products (발효유제품에서 박테리오파지의 특성, 검출과 제어)

  • Ahn, Sung-Il;Azzouny, Rehab A.;Huyen, Tran Thi Thanh;Kwak, Hae-Soo
    • Food Science of Animal Resources
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    • v.29 no.1
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    • pp.1-14
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    • 2009
  • This study was to review the classification, detection and control of bacteriophage in fermented dairy products. Bacteriophage has lytic and/or lysogenic life cycles. Epidemiologically speaking, detected major phages are c2, 936 and p335. Among them p335 has been the largest concern in dairy industry. Traditionally, various analytical technologies, such as spot, starter activity, indicator test, ATP measurement and conductimetric analysis, have been used for the phage detection. In recent years, advanced methods such as flow cytometric method, petrifilm, enzyme linked immunosorbent assay (ELISA) and multiflex PCR diagnostic kit have been deveoloped. The phage contamination has been controlled by using heat, high-pressure treatment, and the combinations of heat and pressure, and/or chemical. Also some starter cultures with phage-resistant character have been developed to minimize the concentration of phages in dairy product. Bacteriophage inhibition media such as calcium medium was also mentioned. To prevent the contamination of bacteriophage in dairy industry, further researches on the detection and control of phage, and phage resistant starters are necessary in the future.