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J Immunol Methods ; 492: 112996, 2021 05.
Artículo en Inglés | MEDLINE | ID: covidwho-1078017

RESUMEN

Dried blood spots (DBS) are routinely used in screening newborns for treatable disorders. Immunoglobulin extraction from DBS, serum or other biological fluids loaded on filter paper cards could represent a valuable method of specimen preservation in monitoring immune response against pathogens as well as vaccination efficiency. In this study using different sources including serum, and monoclonal antibodies we established parameters for antibody extraction from the filter cards to assess antibody reactivity against Helicobacter pylori, measles virus (MV) and the novel coronavirus SARS-CoV-2 antigens. We demonstrated that DBS and dried undiluted serum result in completely preserved antibody activity for immunoassays, including in virus neutralization assays against MV. Extraction efficiency was determined by IgG concentration measurements. The plaque-reduction neutralization titer 50% of dried human serum spots remained stable after more than 10-day storage - 1:359 vs. 1:345 for the corresponding frozen sample. DBSs could be used to monitor immune response to bacterial and viral antigens following natural exposure or immunization. Mice immunized with recombinant spike protein receptor-binding domain of SARS-CoV-2 developed a strong antibody response by day 14 and reached titers above 1:64,000 on day 21 following the secondary boost immunization as measured on DBS samples in antigen-mediated ELISA. Variability in IgG concentration of eluted DBS could be influenced by factors involved in sample application, extraction process and sample characteristics. Adjustment of antibody specific activity to the eluted IgG concentration can increase accuracy of the result interpretation, including in SARS-CoV-2 serological diagnostics.


Asunto(s)
Anticuerpos Antivirales/aislamiento & purificación , COVID-19/diagnóstico , Pruebas con Sangre Seca , Infecciones por Helicobacter/diagnóstico , Helicobacter pylori/fisiología , Inmunoensayo/métodos , Virus del Sarampión/fisiología , Sarampión/diagnóstico , Monitorización Inmunológica/métodos , SARS-CoV-2/fisiología , Animales , Anticuerpos Monoclonales , Formación de Anticuerpos , Pruebas con Sangre Seca/métodos , Femenino , Humanos , Inmunidad Humoral , Ratones , Ratones Endogámicos BALB C , Pruebas Serológicas , Glicoproteína de la Espiga del Coronavirus/inmunología , Vacunación
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