High sensitivity enzyme-linked immunosorbent assay (ELISA) method for measuring protein carbonyl in samples with low amounts of protein

2005 ◽  
Vol 39 (10) ◽  
pp. 1362-1367 ◽  
Author(s):  
D ALAMDARI ◽  
E KOSTIDOU ◽  
K PALETAS ◽  
M SARIGIANNI ◽  
A KONSTAS ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
High Sensitivity ◽  
Protein Carbonyl ◽  
Enzyme Linked Immunosorbent Assay ◽  
Elisa Method

scholarly journals Interpretations of Antibody Responses toSalmonella enterica Serotype Enteritidis gm Flagellin in Poultry Flocks Are Enhanced by a Kinetics-Based Enzyme-Linked Immunosorbent Assay

1998 ◽  
Vol 5 (4) ◽  
pp. 550-555 ◽  
Author(s):  
Patrick L. McDonough ◽  
Richard H. Jacobson ◽  
John F. Timoney ◽  
Ahmed Mutalib ◽  
David C. Kradel ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
High Sensitivity ◽  
High Specificity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Analytical Sensitivity ◽  
Department Of Agriculture ◽  
Trace Back ◽  
Elisa Format ◽  
Commercial Poultry ◽  
Computer Controlled

ABSTRACT Many regulatory and diagnostic programs for the detection ofSalmonella enterica serotype Enteritidis infection in commercial poultry flocks have relied on rapid Pullorum agglutination tests to screen birds because of the shared antigens of S. enterica Enteritidis and S. enterica Pullorum and Gallinarum; however, the use of the enzyme-linked immunosorbent assay (ELISA) format affords better analytical sensitivity than crude agglutination tests. In this study, we adapted our earlier conventional indirect ELISA, using gm flagellin as the antigen, to a kinetics-based, computer-controlled ELISA (KELA). The KELA was used to screen for flagellin antibody from three commercial flocks: (i) a large flock involved in a U.S. Department of Agriculture trace back from a humanS. enterica Enteritidis foodborne outbreak (n = 3,209), (ii) a flock infected with the endemicS. enterica Enteritidis serotype but which also had multiple other salmonella serotypes (n = 65), and (iii) an S. enterica Pullorum-infected flock (n = 12). The first flock (S. entericaEnteritidis prevalence of 2.45% based on culture) provided a field test of the KELA and allowed the calculation of diagnostic sensitivity (D-Sn) and diagnostic specificity (D-Sp). With a cutoff of 10 (used for screening flocks [i.e., high sensitivity]), the KELA has a D-Sn of 95.2% and a D-Sp of 18.5%; with a cutoff of 140 (used in confirmatory flock testing [i.e., high specificity]), the KELA has a D-Sn of 28.0% and a D-Sp of 99.1%. We found that with a cutoff of 60 (D-Sn = 63.1%; D-Sp = 91.6%), we could eliminate reactions in the KELA caused by other non-S. enterica Enteritidis salmonellae. The KELA was also compared to two commercial rapid Pullorum tests, the Solvay (D-Sn = 94.9%; D-Sp = 55.5%) and the Vineland (D-Sn = 62.0%; D-Sp = 75.3%).

An enzyme-linked immunosorbent assay for the detection of apple mosaic virus in yellow birch

1980 ◽  
Vol 10 (3) ◽  
pp. 278-283 ◽  
Author(s):  
T. Hardcastle ◽  
A. R. Gotlieb
Keyword(s):  
Mosaic Virus ◽  
Immunosorbent Assay ◽  
Leaf Tissue ◽  
Enzyme Linked Immunosorbent Assay ◽  
Elisa Method ◽  
Yellow Birch ◽  
Field Samples

An enzyme-linked immunosorbent assay (ELISA) method was developed to detect apple mosaic virus (ApMV) in Betulaalleghaniensis Britton. Tests for virus using bark and bud tissues of dormant trees were successful. ApMV was also detectable in old leaf tissue in August and September, as well as in newly emerging leaf tissue forced in a greenhouse in March. Whole crude antiserum used to coat ELISA plates in tests with bud tissues was a reliable substitute for purified immunoglobulin without loss of sensitivity or specificity. An attempt was made to use ELISA for quantifying virus concentrations in field samples of ApMV-infected birch leaves.

scholarly journals Development of an Enzyme-Linked Immunosorbent Assay for Serotyping Ureaplasma urealyticum Strains Using Monoclonal Antibodies

2001 ◽  
Vol 8 (1) ◽  
pp. 52-57 ◽  
Author(s):  
Fedoua Echahidi ◽  
Gaëtan Muyldermans ◽  
Sabine Lauwers ◽  
Anne Naessens
Keyword(s):  
Monoclonal Antibodies ◽  
Immunosorbent Assay ◽  
Ureaplasma Urealyticum ◽  
Polyclonal Antibodies ◽  
Enzyme Linked Immunosorbent Assay ◽  
Elisa Method ◽  
Cross Reactions ◽  
Antigenic Preparation ◽  
Negative Controls ◽  
Reference Strains

ABSTRACT Ureaplasma urealyticum comprises 14 serotypes. The existing serotyping methods all use polyclonal antibodies. These methods are time-consuming and labor-intensive, and they cannot always be performed on primary isolates; in addition, the results are difficult to interpret. We developed a new enzyme-linked immunosorbent assay (ELISA) method using serotype-specific monoclonal antibodies (MAbs) to enable the serotyping of U. urealyticum isolates from primary broth cultures. Each of the 14 serotype reference strains was tested against 14 selected MAbs. Homologous reactions were very strong, while heterologous reactions were negligible. Three cross-reactions were observed: MAb 5 cross-reacted with serotype 2, MAb 14 cross-reacted with serotype 3, and MAb 8 cross-reacted with serotype 13. Despite the cross-reactions observed, all the serotype reference strains ofU. urealyticum could be identified and differentiated using a combination of MAbs. Reproducibility was analyzed with a fractionated antigenic preparation and with several freshly prepared antigens of the same strain. No significant interrun variation was found with the fractionated antigen, but significant variations in optical density (OD) values were found when freshly prepared antigens were tested. However, the variation in OD values did not influence the overall interpretation of the ELISA: reactions with homologous MAbs were always prominent compared to those of the negative controls. This newly developed ELISA using MAbs seems promising for serotyping of U. urealyticum clinical isolates.

scholarly journals High Sensitivity C-Reactive Protein sebagai Parameter Diagnostik dan Prediktor Luaran Sepsis pada Anak yang Menderita Systemic Inflammatory Response Syndrome

Sari Pediatri ◽  
2016 ◽  
Vol 16 (4) ◽  
pp. 278
Author(s):  
Sofni Sarmen ◽  
Mayetti Mayetti ◽  
Hafni Bachtiar
Keyword(s):  
Systemic Inflammatory Response Syndrome ◽  
Inflammatory Response ◽  
Immunosorbent Assay ◽  
High Sensitivity ◽  
Systemic Inflammatory Response ◽  
Enzyme Linked Immunosorbent Assay ◽  
C Reactive Protein ◽  
Reactive Protein ◽  
Hs Crp ◽  
Elisa Data

Latar belakang. Sepsis merupakan salah satu penyebab morbiditas dan mortalitas pada anak. Diagnosissepsis ditegakkan berdasarkan gejala Systemic Inflammatory Response Syndrome (SIRS) dan penemuan bakteripada kultur darah. Kultur bakteri darah memiliki sensitifitas yang rendah dan membutuhkan waktu yanglama sehingga sering menyebabkan terjadinya overdiagnosis dan overtreatment. C-reactive protein adalahreaktan fase akut yang kadarnya meningkat pada keadaan infeksi. High sensitivity C-reactive protein (hs-CRP) adalah metode yang lebih sensitif untuk mengukur kadar CRP dalam jumlah kecil.Tujuan. Mengetahui peran hs-CRP sebagai parameter diagnostik dan prediktor luaran sepsis pada anakyang menderita SIRS.Metode. Penelitian uji diagnostik dengan desain potong lintang terhadap 85 anak dengan gejala SIRS berusia1 bulan sampai dengan 15 tahun dan dirawat di bangsal anak RS.Dr.M.Djamil Padang sejak Juni sampaiNovember 2012. Pemeriksaan hs-CRP dilakukan dengan metode enzyme-linked immunosorbent assay (ELISA).Data dianalisis dengan SPSS serta dilakukan uji diagnostik. Baku emas sepsis adalah biakan darah.Hasil. Cut off point hs-CRP untuk menentukan sepsis adalah 15,55 ng/ml, (sensitivitas 90,9% dan spesivisitas53,8%). Kadar rata-rata hs-CRP meningkat sesuai dengan beratnya penyakit.Kesimpulan. High sensitivity C-reactive protein dapat dijadikan sebagai parameter diagnostik sepsis padapasien SIRS dengan cut off point 15,55 ng/ml, serta dapat dipakai sebagai prediktor luaran sepsis.

scholarly journals SOME RESULTS OF RESEARCH WORK ON PREVENTION OF ECHINOCOCCOSIS IN KYRGYZSTAN

2020 ◽  
pp. 428-431
Author(s):  
Tursunov ◽  
Isaev ◽  
Ibragimova
Keyword(s):  
Veterinary Medicine ◽  
Immunosorbent Assay ◽  
Research Work ◽  
Preventive Treatment ◽  
Enzyme Linked Immunosorbent Assay ◽  
Positive Outcomes ◽  
Kyrgyz Republic ◽  
Laboratory Studies ◽  
Elisa Method ◽  
Almost All

The Kyrgyz Research Institute of Veterinary Medicine is a co-executor of the Strategy for the Echinococcosis Control (2013–2018) and monitored dogs epizootological examinations after preventive treatment with azinox. Laboratory studies of biomaterials (feces) for infection with helminths were carried out along with the traditional coprological method, a more sensitive enzyme-linked immunosorbent assay (ELISA). The results of laboratory studies of fecal samples from dogs showed that the activities of the "Strategy for the Control of Echinococcosis in the Kyrgyz Republic", including the treatment of dogs with azinox, gave positive outcomes. From 2014 to 2018 2260 feces samples from deworming dogs from almost all regions of Kyrgyzstan were examined. At the same time, the infection of dogs with taeniidae is steadily decreasing; it has also been noted that the main joints of parasitocenosis in dogs are taeniidae, toxocaras, mesocestoids, and eimeria. In the studies conducted on the infection of dogs with taeniidae, the coprological and more sensitive ELISA method was used. Using both helminthological studies, a decrease in the invasion of dogs by taeniidae was confirmed. Thus, in the period from 2014 (the beginning of the Strategy) to 2018, the average invasion of dogs was reduced from 20.2 to 14.3%.

Diarrhetic Shellfish Poisoning: Evaluation of Enzyme-Linked Immunosorbent Assay Methods for Determination of Dinophyslstoxin-2

1995 ◽  
Vol 78 (6) ◽  
pp. 1403-1407 ◽  
Author(s):  
Eoin P Carmody ◽  
Kevin J James ◽  
Seán S Kelly
Keyword(s):  
Immunosorbent Assay ◽  
Cross Reactivity ◽  
Regulatory Control ◽  
Enzyme Linked Immunosorbent Assay ◽  
Shellfish Poisoning ◽  
Diarrhetic Shellfish Poisoning ◽  
Elisa Method ◽  
Multiple Samples ◽  
Liquid Chromatographic

Abstract Dinophysistoxin-2 (DTX-2), an isomer of okadaic acid (OA), recently has been found in Irish waters. DTX-2 was the predominant toxin during prolonged infestations in cultivated mussels along the southwest coast of Ireland. Substantial variations in toxin levels may exist both horizontally and vertically in the water column. The need to take multiple samples and the ethical concern about the use of mammals for routine quality control of shellfish prompted examination of 2 commercially available enzyme-linked immunosorbent assay (ELISA) methods, designed to detect OA, for determination of both OA and DTX-2. One ELISA method (DSPCheck, Sceti Co. Ltd., (Tokyo, Japan) showed good cross-reactivity (40 ± 5%) with standard DTX-2. This study showed that both ELISA methods show good correlation with the liquid chromatographic analysis of 9-anthryldiazomethane derivatives when OA is the predominant toxin present. The sensitivity was also good for OA determination using both methods, which allowed toxin measurement at 10 ng/mL (0.5 ng/well). This level is equivalent to 0.03 μg/g mussel meat. Blank mussel samples spiked with DTX-2 standards gave a good linear correlation (r = 0.997) with this ELISA method when toxin levels were 0.03-0.3 μg/g mussel meat. This range is appropriate for regulatory control of diarrhetic shellfish poisoning.

scholarly journals Evaluation of Solid-Phase Chemiluminescent Enzyme Immunoassay, Enzyme-Linked Immunosorbent Assay, and Latex Agglutination Tests for Screening Toxoplasma IgG in Samples Obtained from Cats and Pigs

2000 ◽  
Vol 12 (2) ◽  
pp. 136-141 ◽  
Author(s):  
Ashok K. Singh
Keyword(s):  
Enzyme Immunoassay ◽  
Immunosorbent Assay ◽  
Solid Phase ◽  
Latex Agglutination ◽  
Clinical Samples ◽  
Enzyme Linked Immunosorbent Assay ◽  
Elisa Method ◽  
Serum Samples ◽  
Positive Results ◽  
Chemiluminescent Enzyme Immunoassay

Serum samples from cats and pigs were analyzed by the solid-phase chemiluminescent enzyme immunoassay (SPCEI), enzyme-linked immunosorbent assay (ELISA), and indirect latex agglutination (ILA) methods. The SPCEI and ILA methods accurately analyzed Toxoplasma IgG (T-IgG) in both clinical and spiked samples from pigs and cats. The ELISA method accurately analyzed T-IgG in spiked samples from cats and pigs or clinical samples from pigs, but it did not accurately analyze T-IgG in clinical samples from cats. The antibody used in the ELISA kit did not cross-react with cat T-IgG. The SPCEI method that uses a stand-alone automated analyzer provided quantitative analysis, whereas the ELISA and ILA methods provided qualitative or, at best, semiquantitative analysis of T-IgG. The SPCEI and ELISA methods were rapid (60–90 minutes for 30 samples), whereas the ILA method required 13–15 hours for 30 samples. Although the three methods accurately distinguished positive from negative samples, the ILA method yielded many weakly positive results that were not confirmed by either the ELISA or SPCEI method. Thus, the indirect agglutination tests may give nonspecific responses at lower T-IgG concentrations.

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