Multiplexed Bead Immunoassays: Advantages and Limitations in Pediatrics

Burgos‐Ramos, Emma; Martos‐Moreno, Gabriel Ángel; Argente, Jesús; Barrios, Vicente

doi:10.5772/36024

Cited by 4 publications

(5 citation statements)

References 36 publications

(30 reference statements)

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“…Multiplexed bead-based immunoassays (MBIA) offer an efficient option of multiplexing strategy, in which sets of magnetic microspheres coated with different peptides of interest offer a platform for the detection of antibodies to different, structurally relevant pathogens or to different, antigenic targets of a single pathogen or both, all in a single procedure [8].…”

Section: Introductionmentioning

confidence: 99%

Characterization of Immune Responses to SARS-CoV-2 and Other Human Pathogenic Coronaviruses Using a Multiplex Bead-Based Immunoassay

et al. 2021

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Serological assays that simultaneously detect antibodies to multiple targets of SARS-CoV-2 and to other structurally related coronaviruses provide a holistic picture of antibody response patterns. Well-validated multiplex immunoassays are scarce. Here, we evaluated the performance of an 11-plex serological assay capable of detecting antibodies directed to four antigenic targets of SARS-CoV-2 and to S1 proteins of other human pathogenic coronaviruses. We used 620 well-characterized sera (n = 458 seropositive and n = 110 seronegative for SARS-CoV-2 in the pre-SARS-CoV-2 era and n = 52 seronegative for SARS-CoV-2 in the era of SARS-CoV-2) as positive and negative standards. We calculated the sensitivity, specificity, as well as positive and negative predictive values, including a 95% confidence interval. The difference in mean fluorescence intensity (95% CI) was used to assess a potential cross-reaction between antibodies to SARS-CoV-2 and the other coronaviruses. The sensitivity (95% CI) of detecting anti-SARS-CoV-2 antibodies to four antigenic targets ranged from 83.4% (76.7–86.7) to 93.7% (91.0–95.7) and the specificity from 98.2% (93.6–99.8) to 100% (96.7–100). We observed no obvious cross-reaction between anti-SARS-CoV-2 antibodies and antibodies to the other coronaviruses except for SARS-CoV-1. The high sensitivity and specificity warrant a reliable utilization of the assay in population-based seroprevalence surveys or vaccine efficacy studies.

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Section: Introductionmentioning

confidence: 99%

Characterization of Immune Responses to SARS-CoV-2 and Other Human Pathogenic Coronaviruses Using a Multiplex Bead-Based Immunoassay

et al. 2021

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“…Using the AMIA, the AFP/PSA/CEA levels in 10 serum samples from six healthy persons and four liver cancer patients are simultaneously measured. Notably, only 20 μL of serum is enough for the AMIA test, highlighting its minimal sample consumption, which is of great importance to practical clinical applications, for instance, in obstetrics and pediatrics. , Meanwhile, we also calculate the testing costs per AMIA assay (Table S3); with only $6, three kinds of protein biomarkers can be simultaneously detected. The target antigens in the same batch of serum samples are also clinically assessed using an ADVIA Centaur CP chemiluminescence platform (SIEMENS, Germany) from the Mengchao Hepatobiliary Hospital of Fujian Medical University.…”

Section: Resultsmentioning

confidence: 99%

Integrated Single Microbead-Arrayed μ-Fluidic Platform for the Automated Detection of Multiplexed Biomarkers

et al. 2020

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An automated, single microbead-arrayed μ-fluidic immunoassay (AMIA) device is innovatively devised in this study, which enables the highly sensitive and simultaneous detection of multiplex biomarkers with fully automatic operations. The AMIA platform not only achieves automated assay processing and multiplexed target detection by integrating single microbead manipulation, sample loading, multistep washing, and immunoreaction on a microfluidic chip but also confers high sensitivity due to the highly efficient signal enriching effect on a single microbead by the use of only a routine sandwich immunoreaction. As such, as low as the pg/mL level of multiplexed protein biomarkers can be simultaneously determined in a quite small volume of serum (∼20 μL is enough), which can well meet the clinical demand for disease screening and prognosis. What is more, the detection results of several clinically important biomarkers in clinical samples with the AMIA platform exhibit excellent consistency with those obtained by using a standard clinical test. Thus, in virtue of the excellent features in terms of high sensitivity, multiplexing capability, generality, and high degree of automation, the AMIA provides a practical and user-friendly platform for assaying different biomarkers in clinical diagnostics and point-of-care testing.

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“…Magnetic ELISA also provides the availability of certain epitopes on the surface of the granules, that is, the availability of primary antibodies, in the case of setting sandwich variants of the method (Kourilov et al, 2002). The limitation in the formulation of a magnetic ELISA still is the interpretation of the results obtained at very low concentrations of target substances in the samples (Burgos-Ramos et al, 2012).…”

Section: Comparative Characteristics Of Mia and Its Prospectsmentioning

confidence: 99%

“…Another application of MIA is the detection of mutations in medical genetics. This method has been used for genotyping in samples from patients with predisposition to thrombophilia and detecting fusion transcripts of chromosomal translocations in children with acute lymphoblastic leukemia (Burgos-Ramos et al, 2012). MIA can also be used to detect pathogens and toxins present in food, water analysis (Orlov et al, 2013).…”

Section: Comparative Characteristics Of Mia and Its Prospectsmentioning

confidence: 99%

Modern magnetic immunoassay: Biophysical and biochemical aspects

Galkin

Besarab

Pysmenna

et al. 2017

Regul. Mech. Biosyst.

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In this review article an analysis of the biochemical and biophysical aspects of modern magnetic immunoassay (MIA) is conducted and additionally the problems and perspectives of its application in biology, biotechnology and medicine are defined. Magnetic immunoassay should be considered as an evolutionary extension of the classical immunoassay. MIA can have many variants of modifications, similar to the classic immunoenzymatic assay. The key distinctive element of the MIA is the use of magnetic particles (MPs), which are usually nanoparticles. MPs in the MIA can act as a marker for detection, or the solid phase at which the immunochemical reaction takes place. MIA possesses basic advantages over classical immunoassay methods: thanks to the unique magnetic properties of the MPs and the ability to manipulate it in the external magnetic field, it is possible to increase the informative value of the analysis (first of all, sensitivity and specificity), as well as the rigid requirements for “purity” of tested samples. For the purposes of immunoassay, magnetic particles of size from 10 to 200 nm are important, since such particles are in a superparamagnetic state, in the absence of strong magnetic fields; they are not agglomerated in a liquid medium. The size of the spherical particle determines the rate of sedimentation and mobility in the solution. The outer polymeric membrane serves as a matrix in which the surface functional groups are added, and also protects the core of the metal from the external environment. The outer shell may also consist of agarose, cellulose, porous glass, silicon dioxide etc. There are several strategies for the synthesis of nanoparticles: mechanical (dispersion), physical (gas phase deposition), wet chemical methods (chemical comprecipitation, thermal decomposition, methods of micro emulsion, hydrothermal reactions) and physico-chemical methods. Also used are magnesite nanoparticles of biogenic origin. Magnetic particles may function, and this is important for immunoassay. Surface functional groups include carboxylic, amino, epoxy, hydroxyl, tosyl, and N-hydroxysuccinate-activated groups. Magnetic spherical particles usually interact with surface molecules such as streptovidine, biotin, protein A, protein G, and immunoglobulin etc. Directions and prospects of the development of methods of magnetic immunoassay are determined, mainly, by the development of methods for detecting or influencing magnetic particles. In this case, the classical methods of detection are electrochemical methods, electrochemiluminescence, fluorescence. More modern ones include giant magnetoresistance, superconducting quantum interference devices, surface-enhanced Raman spectroscopy, biosensors based on nonlinear magnetization, magneto-PCR immunoassay. The current trend is to combine or integrate the application of various biochemical, physical, molecular and genetic, physico-chemical detection methods. In fact, all of these benefits undoubtedly open up broad prospects for the practical application of MIA in biology, biotechnology and medicine.

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Multiplexed Bead Immunoassays: Advantages and Limitations in Pediatrics

Cited by 4 publications

References 36 publications

Characterization of Immune Responses to SARS-CoV-2 and Other Human Pathogenic Coronaviruses Using a Multiplex Bead-Based Immunoassay

Characterization of Immune Responses to SARS-CoV-2 and Other Human Pathogenic Coronaviruses Using a Multiplex Bead-Based Immunoassay

Integrated Single Microbead-Arrayed μ-Fluidic Platform for the Automated Detection of Multiplexed Biomarkers

Modern magnetic immunoassay: Biophysical and biochemical aspects

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