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1.
Methods Mol Biol ; 2511: 257-271, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35838966

RESUMO

Serological assays have been a useful tool for detection of antibodies to SARS-CoV-2 during the COVID-19 pandemic. These assays are used for epidemiology and serosurveillance to monitor the progression of the pandemic, to identify and differentiate individuals who have developed antibodies from natural infection versus vaccine-induced immunity, and to identify potential donors of convalescent plasma for therapeutic purposes. In this chapter, we describe a commercially available bead-based serological assay, the Luminex® xMAP® SARS-CoV-2 Multi-Antigen IgG Assay, that detects and identifies antibodies against three SARS-CoV-2 antigens. In addition to the assay principle and workflow, we describe modifications that may be used to evaluate alternate sample types, antibody isotypes, and potential neutralizing antibody responses.


Assuntos
COVID-19 , SARS-CoV-2 , Anticorpos Neutralizantes , Anticorpos Antivirais , COVID-19/diagnóstico , COVID-19/terapia , Humanos , Imunização Passiva , Imunoglobulina G , Pandemias , Sensibilidade e Especificidade , Soroterapia para COVID-19
2.
Alzheimers Dement (Amst) ; 6: 201-209, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28349119

RESUMO

INTRODUCTION: Changes in cerebrospinal fluid (CSF) tau and amyloid ß (Aß)42 accompany development of Alzheimer's brain pathology. Robust tau and Aß42 immunoassays were developed to establish a tau/Aß42 cutoff distinguishing mild-to-moderate Alzheimer's disease (AD) subjects from healthy elderly control (HC) subjects. METHODS: A CSF tau/Aß42 cutoff criteria was chosen, which distinguished the groups and maximized concordance with amyloid PET. Performance was assessed using an independent validation cohort. RESULTS: A tau/Aß42 = 0.215 cutoff provided 94.8% sensitivity and 77.7% specificity. Concordance with PET visual reads was estimated at 86.9% in a ∼50% PET positive population. In the validation cohort, the cutoff demonstrated 78.4% sensitivity and 84.9% specificity to distinguish the AD and HC populations. DISCUSSION: A tau/Aß42 cutoff with acceptable sensitivity and specificity distinguished HC from mild-to-moderate AD subjects and maximized concordance to brain amyloidosis. The defined cutoff demonstrated that CSF analysis may be useful as a surrogate to imaging assessment of AD pathology.

3.
Cytometry A ; 69(5): 384-90, 2006 May.
Artigo em Inglês | MEDLINE | ID: mdl-16615139

RESUMO

INTRODUCTION: Typically, bead-based assays ("bead arrays") use the mean or median value of a population of measurements to judge ligand binding or other activity, which results in a change in fluorescence intensity. Individual bead measurements are used here to calculate population parameters integral to the measurement of a bead array. METHODS: Using a commercially-available instrument designed for bead array measurements, a set of beads were labeled with biotin and then titrated with PE-Streptavidin. Data were collected under normal machine conditions as well as variations. RESULTS: The "sensitivity" of the measurements was determined using parametric and nonparametric statistical methods as well as regression analysis over a limited range of the titration (concentration vs. response profile). CONCLUSIONS: Results at low ligand concentrations suggest that precise measurements with bead array systems require a large number of individual bead measurements to be acquired. Individual bead measurements should be used to determine the mean and confidence intervals for the calculated measurements. These results also apply to regression analysis of concentration-response profiles. Furthermore, features of the instrument can be manipulated to achieve increased sensitivity and detection of lower amounts of ligand bound to the bead populations.


Assuntos
Fluorometria/instrumentação , Microesferas , Biotina/química , Biotina/metabolismo , Fluorescência , Análise de Regressão , Sensibilidade e Especificidade , Estreptavidina/química , Estreptavidina/metabolismo
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