• Title/Summary/Keyword: ELISA kit

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Comparison of an Immunochromatographic Assay Kit with DAS-ELISA for Large-Scale Diagnosis and Molecular Discrimination of Satsuma Dwarf Virus Collected from Citrus Orchards

  • Kato, Mitsuhiro;Tomimura, Kenta;Ishii, Kanako
    • The Plant Pathology Journal
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    • v.36 no.5
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    • pp.509-514
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    • 2020
  • Satsuma dwarf virus (SDV) seriously damages citrus production by reducing the quality and yield of fruit. To avoid contamination with SDV, mother trees are checked to be SDV-free in advance of nursery tree distribution. In this study, we compared an immunochromatographic assay (ICA) kit with double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) for large-scale diagnosis of SDV in orchardgrown trees in Shizuoka Prefecture, Japan. The two methods gave conflicting results for 11 of 1,705 samples, all of which were negative by DAS-ELISA but positive by ICA. The samples scored as positive by either DAS-ELISA or ICA were analyzed by reverse transcription polymerase chain reaction and all were confirmed to be positive. These results validate the use of ICA as a screening method for large-scale diagnosis. Strain discrimination revealed that 16 of 22 isolates belonged to SDV, while citrus mosaic virus (CiMV) infection only and co-infection (SDV and CiMV) were in a minority.

A New ELISA Kit Based on Antigenic Epitopes for Diagnosing Brucella abortus

  • Khodabakhsh, Tannaz;Arabi, Azin;Pakzad, Parviz;Gheflat, ShivaSadat;Bahreinipour, Akram;Bandehpour, Mojgan
    • Microbiology and Biotechnology Letters
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    • v.47 no.1
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    • pp.158-163
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    • 2019
  • Brucellosis is one of the most important zoonotic diseases that lead to a great amount of economic losses. Prevention and diagnosis are both necessary to eradicate this disease. The identification and evaluation of different antigens of Brucella spp. play a key role in the progress of diagnostic programs. In this study, we designed, produced, and evaluated a 24-kDa polypeptide containing antigenic epitopes of VirB2, 3, and 9 of Brucella abortus for use with the ELISA kit. The produced polypeptide is appropriate for diagnosing brucellosis in bovines by a laboratory diagnostic kit, with 100% sensitivity and 97.5% specificity.

Nested-PCR and a New ELISA-Based NovaLisa Test Kit for Malaria Diagnosis in an Endemic Area of Thailand

  • Thongdee, Pimwan;Chaijaroenkul, Wanna;Kuesap, Jiraporn;Na-Bangchang, Kesara
    • Parasites, Hosts and Diseases
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    • v.52 no.4
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    • pp.377-381
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    • 2014
  • Microscopy is considered as the gold standard for malaria diagnosis although its wide application is limited by the requirement of highly experienced microscopists. PCR and serological tests provide efficient diagnostic performance and have been applied for malaria diagnosis and research. The aim of this study was to investigate the diagnostic performance of nested PCR and a recently developed an ELISA-based new rapid diagnosis test (RDT), NovaLisa test kit, for diagnosis of malaria infection, using microscopic method as the gold standard. The performance of nested-PCR as a malaria diagnostic tool is excellent with respect to its high accuracy, sensitivity, specificity, and ability to discriminate Plasmodium species. The sensitivity and specificity of nested-PCR compared with the microscopic method for detection of Plasmodium falciparum, Plasmodium vivax, and P. falciparum/P. vivax mixed infection were 71.4 vs 100%, 100 vs 98.7%, and 100 vs 95.0%, respectively. The sensitivity and specificity of the ELISA-based NovaLisa test kit compared with the microscopic method for detection of Plasmodium genus were 89.0 vs 91.6%, respectively. NovaLisa test kit provided comparable diagnostic performance. Its relatively low cost, simplicity, and rapidity enables large scale field application.

A study on production of early pregnancy diagnostic kit in cattle II. Production of early pregnancy diagnostic kit (소의 조기 임신진단 kit의 개발 II. 조기 임신진단 kit의 개발)

  • Kang, Chung-boo;Lee, Hyo-jong;Choe, Sang-yong
    • Korean Journal of Veterinary Research
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    • v.31 no.2
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    • pp.223-228
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    • 1991
  • Most progesterone enzyme immunoassays(EIA) are used liquid-phase double-antibody or single-antibody seperation. These methods consume considerable time and reagents because of the requirements for several washing and centrifugation steps involving the reactants. Because of these several problems, we were prompted to develop an effective enzyme-linked immunosorbent assay(ELISA) system that would be equal or superior to RIA for assay of progesterone. The results were obtained as follows. 1. Cross reaction of the progesterone antiserum with other steroids determined was shown with progesterone(100%), $11{\alpha}$-deoxycorti-costerone(2.271%), but the other steroids were shown below 0.9%. 2. Standard curve for progesterone ELISA was shown available difference according to progesterone concentration from 0 to 1,000pg/ml. 3. The lower limit of sensitivity was 0.2pg/well 4. Progesterone concentration was 1.6ng/ml for before parturition, and that was below 0.5ng/ml for after parturition. This development enzyme-linked immunosorbent assay for progesterone can be detected pregnancy diagnosis in cattle, and also applicable 10 research on physiological function including such as reproductive disorders.

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Development of Infectious Bronchitis Virus (IBV) ELISA Kit for Detection of Antibodies against Nephropathogenic IBV Vaccine (국내회사와 다국적기업 제조 ELISA 키트의 전염성 기관지염 백신에 따른 항체 검출능 비교)

  • Kim, Kyu-Jik;Kim, Jun-Young;Youn, Ha-Na;Ju, Hyo-Sun;Lee, Da-Yeah;Song, Chang-Seon
    • Korean Journal of Poultry Science
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    • v.45 no.1
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    • pp.17-28
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    • 2018
  • Infectious bronchitis virus (IBV) is an economically important disease in the poultry industry worldwide. This disease commonly manifests respiratory signs, poor egg quality, and decline in egg production. Since IBV is a RNA virus, the emergence of new variant strains is continuously reported and the immunization of susceptible chickens with only one antigenic type of the virus has been shown to induce partial or no protection against other unrelated types. Therefore, it is difficult to diagnose IBV due to variants serotypes. In this study, we collected serum from various ages of Broiler GP (Grandparent) to Layer CC (Commercial chick) and performed detectability comparison test between domestic company and multinational company manufacturing ELISA kit. Results of this experiment suggest that domestic company manufacturing ELISA kit is more sensitive to infectious bronchitis antibody than that of the multinational company. Our findings also suggest that antibody's change trends after infectious bronchitis vaccination. Thus, the use of appropriate kit for domestic situations is important.

Determination of urinary cortisol : creatinine ratios by sequential thin layer chromatography and ELISA in dogs (개에서 thin layer chromatography 박층크로마토그라피 및 ELISA를 이용한 요중의 cortisol : creatinine비 측정)

  • Sohn, Dae-ho;Na, Ki-jeong;Oh, Tae-ho;Lee, Hye-sook;Han, Hong-ryul
    • Korean Journal of Veterinary Research
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    • v.39 no.5
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    • pp.1006-1012
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    • 1999
  • This study was conducted to evaluate the ELISA kit for measuring the level of cortisol in the urine. The CV of within-run variation and day to day variation were 0.4~2.8 and 1.8~5.7, respectively. The minimum limitation of measurement was 1ng/ml. The cross reaction was high ($CR_{50}(%)=11.4{\sim}43.2$) in prednisolone, 11-deoxycortisol, 21-deoxycortisol and predinosone. There was low and no cross reaction in other steroid. To develop the ELISA kit we measured the cortisol level in diluted urine with PBS (procedure I), extracted urine with methylene chloride (procedure II) and extracted methylene chloride-extracted urine from thin-layer chromatography (procedure III). The CV value of procedure I, II, III was 9.4~28.3%, 7.2~8.9% and 2.5~5.7%, respectively. There was significant difference between procedure I with II, and pro-cedure I with III(p < 0.01), but no difference between procedure II with III significantly(p < 0.01). The mean UCCR of urine collected through am 8 to 10 was $9.5{\pm}7.6$(0.14~28.0) in 12-month-old dog(n = 47). In this study we can measure the cortisol level in extracted urine with methylene chloride and sequential thin-layer chromatography accurately using ELISA kit.

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Nonspecific Mouse Hepatitis Virus Positivity of Genetically Engineered Mice Determined by ELISA

  • Han, Dae Jong;Kim, Hyuncheol;Yeom, Su-Cheong
    • Biomedical Science Letters
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    • v.21 no.1
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    • pp.9-14
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    • 2015
  • Mouse hepatitis virus (MHV) is a major pathogen in laboratory mice that usually leads to fatal diseases, such as hepatitis, multiple sclerosis, encephalitis, and respiratory disease. MHV has a high infection rate, and it needs to be detected as soon as possible to prevent its spread to other facilities. However, MHV detection by enzyme-linked immunosorbent assay (ELISA) often gives false positives; thus, it is very important that the results are confirmed as true positives in the early infection stage or distinguished as false positives with more accurate, reliable methods. Under microbiological screening, MHV ELISA-positive mice were found in four GFP-tagging transgenic mice. To verify the detection of the MHV antigen directly, reverse transcription polymerase chain reaction (RT-PCR) was performed, and the mice were determined to be MHV negative. Additional serum antibody-based screening was conducted with three different ELISA kits, and multiplexed fluorometric immunoassay (MFIA) was performed to confirm their accuracy/sensitivity. In brief, the ELISA kit for A59 nucleocapsid protein (MHV-A59N) revealed MHV ELISA positivity, while other ELISA kits (MHV-S lysate and MHV-JHM lysate) demonstrated MHV negativity. In MFIA, only the test for the recombinant A59 nucleocapsid antigen was MHV positive, which was consistent with the ELISA results. These results suggest that the ELISA kit with the recombinant A59 nucleocapsid antigen might induce non-specific MHV ELISA positivity and that confirmation is therefore essential.

Survey of Ehrlichia canis and Borrelia burgdorferi antibodies in dogs (German shepherd) reared in Korea

  • Lee, Sang-Eun;Song, Kun-Ho;Lee, Suk-Hoon
    • Korean Journal of Veterinary Research
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    • v.47 no.3
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    • pp.281-283
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    • 2007
  • This study was conducted to survey of Ehrlichia (E.) canis and Borrelia (B.) burgdorferi antibodies among clinically healthy German shepherd dogs (116 females and 120 males) using a ELISA kit (SNAP 3Dx; IDEXX Laboratories, USA) in Korea. Whole blood samples are collected from the 236 dogs and are tested to detect E. canis and B. burgdorferi antibodies by using ELISA kit (SNAP 3Dx; IDEXX Laboratories, USA). Confidence interval comparisions revealed that dogs of 4-6 years have higher prevalence with a seropositive result (CI=0.17-0.45) in E. canis than the other age groups but there are no differences among age groups in B. burgdorferi. Also, no differences with a seropositive result were found among different regions in E. canis and B. burgdorferi antibodies. In conclusion, this study was the first large scale survey of canine E. canis and B. burgdorferi antibodies in Korea and provide an useful reference for clinicians.

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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    • v.51 no.1
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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.

Studies on development of ELISA Kits for T-2 toxin (T-2 독소의 측정을 위한 ELISA Kits의 개발에 관한 연구)

  • Yoon, Hwa-joong;Kim, Taejong;Lee, Sung-yun
    • Korean Journal of Veterinary Research
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    • v.37 no.3
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    • pp.613-618
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    • 1997
  • Direct competitive enzyme linked immunosorbent assay(ELISA) with monoclonal antibodies have been studied for quantitative determination of T-2 toxin from the mold corn. The T-2HS, T-2HS-BSA, T-2HS-HRP and monoclonal antibodies against T-2 toxin produced in the studies were qualified for quantitative ELISA test of T-2 toxin. The mean recovery rate from ground com spiked T-2 toxin was 83%. The meaning range of the T-2 test was 60ng to $2{\mu}g$. According to the recovery results with the com spiked T-2, the tests proved to be suitable in the screening of the moldy feed samples for the presence of T-2 toxin and will be able to become the basis of the ELISA test for the quantitative screening kits of T-2 toxin.

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