RESUMEN
In this work, a quantitative sandwich ELISA was optimized, through a full factorial design of experiments (DOE) in successive steps of a preliminary protocol obtained by the method of one factor at a time (OFAT). The specificity of the optimized ELISA, the lower limit of quantification, the quantification range and the analytical sensitivity of the antigen quantification curve were evaluated, in comparison with the curve obtained from the preliminary protocol. The full factorial DOE was linked to a simple statistical processing, which facilitates the interpretation of the results in those laboratories where there is no trained statistician. The step-by-step optimization of the ELISA and the successive incorporation into the protocol of the best combination of factors and levels, allowed obtaining a specific immunoassay, with an analytical sensitivity 20 times greater and with a lower limit of antigen quantification that decreased from 156.25 at 9.766 ng/mL. As far as we know, there are no reports of optimization of an ELISA following the step-by-step scheme used in this work. The optimized ELISA will be used for the quantification of the TT-P0 protein, the active principle of a vaccine candidate against sea lice.
Asunto(s)
Antígenos , Ensayo de Inmunoadsorción Enzimática/métodos , InmunoensayoRESUMEN
Background: COVID-19 vaccines that trigger a strong secretory antibody response in breast milk may achieve effective passive protection of vulnerable newborns and breastfed infants of immunized mothers. The aim of this work was to investigate the presence of SARS-CoV-2 spike RBD-specific IgA and IgG antibodies in breast milk, 5 and 9 weeks after vaccination with 3 doses of the protein subunit vaccine Abdala, compared to those found in breast milk from COVID-19-recovered women, collected at least 40 days after the infection. Methods: SARS-CoV-2 spike RBD-specific IgA and IgG antibodies were semi-quantified by indirect ELISA, using a homemade standard generated by pooling twenty breast milk samples with high absorbance values according to preliminary data. The validity of the standard curves was proved following the European Medicines Agency Guideline. Two breast milk samples from 2 unvaccinated women who had not been infected with COVID-19 were included as negative controls. Potentially neutralizing antibodies was assessed by a SARS-CoV-2 surrogate virus neutralization test. Results: High levels of anti-RBD IgA antibodies were detected in breast milk samples 9 weeks after vaccination and anti-RBD IgG antibodies rise from the fifth to the ninth week. In the post-COVID-19 time that was evaluated, the IgG-type response was notably higher compared to both post-vaccination periods. Neutralizing antibody titers were similar in breast milk from vaccinated and COVID-19 recovered women. Conclusions: This is the first report about the immune response in breast milk after the administration of a COVID-19 protein subunit vaccine, which could provide analogous protection to that conferred by SARS-CoV-2 infection. This implies a potential passive immunity that breastfed infants receive from their mothers vaccinated with Abdala.
RESUMEN
El presente trabajo de investigación titulado Hipertensión Inducida por el Embarazo y sus consecuencias en el feto y recién nacido en el Hospital Santa Rosa de Puerto Maldonado - Junio 1996 a Junio 1997; es un estudio Retrospectivo, Descriptivo, Transversal. Teniendo como muestra de estudio a 46 gestantes con H.I.E. que cumplieran con los criterios de inclusión y exclusión y sus respectivos recién nacidos, se aplicaron las siguientes pruebas estadísticas: la distribución de fercuencias, media, desviación estándar, intervalo de confianza y chi cuadrado. Se encontró que la mortalidad perinatal fue de 13.04 por ciento más alto que otros estudios nacionales. El S.F.A. presentó un frecuencia del 45 por ciento con alto riesgo en la población expuesta. Se encontró que el 47,82 por ciento eran recién nacidos PEG, el 43,47 por ciento recién nacidos prematuros con un promedio de 36 semanas de gestación y presentaron asfixia neonatal el 65 por ciento al minuto, recuperándose el 61,54 por ciento a los cinco minutos. Se encontró una frecuencia para H.I.E. de 6,03 por ciento para la eclampsia 47,83 por ciento, preeclampsia severa 43,48 por ciento. Los factores de riesgo fueron presión arterial = 140/90 mmHg y PAM = 106 mmHg para ésta población en estudio, la edad de la madre con un promedio de 23 años, la anemia que por una serie de mecanismos lleva a una disminución del flujo útero placentario. Se observa que no es exclusivo de la primera gestación como indica la literatura, si no también se presentó en multíparas con la misma frecuencia que las primíparas.