A comparison of ELISA and flow microsphere-based assays for quantification of immunoglobulins

Dasso, Joseph F.; Lee, Juliet; Bach, Hanh; Mage, Rose G.

doi:10.1016/s0022-1759(02)00028-5

Cited by 69 publications

(43 citation statements)

References 14 publications

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“…The LOD of MIA was similar to ELISA for extracts in which bacteria were detected without previous enrichment. Also in previous comparative studies, the sensitivity of MIA was often similar to ELISA (Dasso et al, 2002;McBride et al, 2003). The LODs of MIA for detection of Pca and Dcd in peel extracts were rather high compared to those published for other bacteria (Dunbar et al, 2003) in which the authors reported a sensitivity of ca.…”

Section: Discussionmentioning

confidence: 61%

“…First, in the MIA multiple beads per sample (100-200) are measured in comparison to the few replicate wells normally used in ELISA per sample. The higher number of replicates result in a higher precision for the MIA (Dasso et al, 2002). Second, the entire MIA can be completed in 1 h, including measurement by the Luminex analyzer which typically takes 20 min per 96-well plate, compared to a minimum of 6 h for ELISA.…”

Section: Discussionmentioning

confidence: 99%

“…Moreover, MIA is less labour-intensive and different analytes can be measured at the same time (Dasso et al, 2002;Biagini et al, 2003). Also when compared to antibody microarrays, the Luminex technology has better features, such as a lower detection limit and a better dynamic range, although microarrays are more suitable for miniaturization (Rao et al, 2004).…”

Section: Introductionmentioning

confidence: 99%

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An enrichment microsphere immunoassay for the detection of Pectobacterium atrosepticum and Dickeya dianthicola in potato tuber extracts

Peters¹,

Śledź

Bergervoet³

et al. 2006

Eur J Plant Pathol

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An enrichment microsphere immunoassay (MIA) was developed, based on the Luminex xMAPÒ technology, for the simultaneous (duplex) detection of Pectobacterium atrosepticum (former name Erwinia carotovora subsp. atroseptica) (Pca) and Dickeya dianthicola (former name Erwinia chrysanthemi) (Dcd) in potato plant extracts. Target bacteria in the extracts were enriched for 48 h in a semi-selective broth containing polypectate under low oxygen conditions. Samples were subsequently incubated with antibodycoated colour-coded microspheres (beads) and with secondary antibodies conjugated with Alexa FluorÒ 532, a reporter dye. Samples were analyzed with the Luminex analyzer, in which one laser identified each microsphere and another laser the reporter dye conjugated to the secondary antibodies. The assay required minimal sample preparation, could be completed in 1 h, was performed in 96 wells microtitreplates and required no wash steps.The limit of detection for the duplex enrichment MIA was 100-1000 cfu ml )1 , which was a hundred times lower than of an enrichment-ELISA. Without enrichment, the sensitivity of MIA and ELISA was largely similar and ranged between 10 6 and 10 7 cells ml )1 . No difference in sensitivity was found between a MIA in a single or duplex format. In a comparative test with non-infected potato plant extracts and extracts from plants infected with Pca or Dcd, results of the enrichment MIA correlated well with those of the enrichment ELISA and enrichment PCR. These results indicate that MIA can be reliably used for multiplex detection of soft rot Enterbacteriaceae in crude potato plant extracts. The technology is an attractive and cost-effective alternative to other detection methods, including ELISA.

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Section: Discussionmentioning

confidence: 61%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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An enrichment microsphere immunoassay for the detection of Pectobacterium atrosepticum and Dickeya dianthicola in potato tuber extracts

Peters¹,

Śledź

Bergervoet³

et al. 2006

Eur J Plant Pathol

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“…Recent availability of commercial instruments based on bead-based immunoassay technology (Dasso et al, 2002;de Jager et al, 2003;Dunbar et al, 2003;Earley et al, 2002;Kellar and Iannone, 2002;Kellar et al, 2001;Prabhakar et al, 2002;Vignali, 2000) is likely to help remove this void. For example, the Bio-Plex™ Suspension Array system (Bio-Rad; Hercules, CA) is an easy-to-use and flexible unit capable of simultaneously analyzing up to 100 (6-150 kDa) proteins from as little as 12 μl of sample, which has been done for analyses of serum (de Jager et al, 2003).…”

Section: Introductionmentioning

confidence: 99%

Optimization of multiplexed bead-based cytokine immunoassays for rat serum and brain tissue

Hulse

Kunkler

Fedynyshyn

et al. 2004

Journal of Neuroscience Methods

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The ability to simultaneously quantify multiple signaling molecule protein levels from microscopic neural tissue samples would be of great benefit to deciphering how they affect brain function. This follows from evidence that indicates signaling molecules can be pleiotropic and can have complex interactive behavior that is regionally and cellularly heterogeneous. Multiplexed examination of tissue proteins has been exceedingly difficult because of the absence of available techniques. This void now has been removed by the commercial availability of bead-based immunoassays for targeted proteins that allow analyses of up to 100 (6-150 kDa) proteins from as little as 12 μl. Thus far used only for sera (human and mouse) and culture media, we demonstrate here that sensitive (as low as 2 pg/ml), wide-ranging (up to 2-32 000 pg/ml), accurate (8% intra-assay covariance) and reliable (4-7% inter-assay covariance) measurements can be made of nine exemplary cytokines (e.g., IL-1α, IL-1β, IL-2, IL-4, IL-6, IL-10, GM-CSF, IFN-γ, TNF-α) simultaneously not only from rat serum but, for the first time, also brain tissue. Furthermore, we describe animal handling procedures that minimize stress as determined by serum glucocorticoid levels since they can influence cytokine expression.

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“…Since the spectrum of molecular entities analyzable this way encompass proteins (1), nucleic acids (2,3), and molecules recognized by these (4-6), the microbead-based flow-cytometric technology can provide a universal measuring platform for many laboratory parameters. Titration of proteins by microbead-based flow-cytometric immunoassays have been demonstrated for several proteins and proved to be viable alternatives to conventional technologies such as ELISAs (7,8). The fields of application include techniques used in microbiology, like immunoassays of bacterial (9) or viral (10) antigens, or analysis of antiviral/ antibacterial antibodies (7,11), both groups of molecules can be a diagnostic aid for such infections.…”

Section: Introductionmentioning

confidence: 99%

Biological microbeads for flow‐cytometric immunoassays, enzyme titrations, and quantitative PCR

Pataki

Szabó²,

Lantos

et al. 2005

Cytometry Pt A

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BackgroundIntroduction of microbeads into flow‐cytometry has created a new scenario, making quantitative measurement of molecules dispersed in a homogeneous phase, with an extremely wide realm of already realized and potential applications possible. Development of this field has lead to specialized instrumentation and microbead arrays, dedicated to certain applications.MethodsFormaldehyde‐fixed yeast and bacterial cells were conjugated with avidin and applied as microbeads, to establish a simple, convenient, flexible, and inexpensive flow‐cytometric platform for various immunological and biochemical assays.ResultsWe have tested these “biological microbeads” for the simultaneous titration of human α‐fetoprotein (AFP) and human Chorionic Gonadotropin (βhCG) hormone levels, for the titration of proteolytic and nucleolytic (restriction) enzymes, and for quantitative PCR, using biotinylated and fluorescent primers.ConclusionsThe use of biological microbeads for various immunological and biochemical assays has been demonstrated. The flow‐cytometric methods proved to be at least as sensitive as the standard biochemical or immunological tests. For proteinase K activity measurements, a single enzyme molecule in the sample could be detected. The sensitivity, versatility, and low cost of the assays may advance flow‐cytometry to become a central methodological platform in most laboratories. The biological microbeads offer virtually unlimited possibilities for fluorescent labeling (addressing), conjugation of ligand binding molecules, and they are easy to handle and perform well in a multiplex format. © 2005 International Society for Analytical Cytology

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A comparison of ELISA and flow microsphere-based assays for quantification of immunoglobulins

Cited by 69 publications

References 14 publications

An enrichment microsphere immunoassay for the detection of Pectobacterium atrosepticum and Dickeya dianthicola in potato tuber extracts

An enrichment microsphere immunoassay for the detection of Pectobacterium atrosepticum and Dickeya dianthicola in potato tuber extracts

Optimization of multiplexed bead-based cytokine immunoassays for rat serum and brain tissue

Biological microbeads for flow‐cytometric immunoassays, enzyme titrations, and quantitative PCR

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