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Mem. Inst. Oswaldo Cruz ; 108(1): 36-40, Feb. 2013. graf, tab
Artículo en Inglés | LILACS | ID: lil-666041

RESUMEN

ELISA in situ can be used to titrate hepatitis A virus (HAV) particles and real-time polymerase chain reaction (RT-PCR) has been shown to be a fast method to quantify the HAV genome. Precise quantification of viral concentration is necessary to distinguish between infectious and non-infectious particles. The purpose of this study was to compare cell culture and RT-PCR quantification results and determine whether HAV genome quantification can be correlated with infectivity. For this purpose, three stocks of undiluted, five-fold diluted and 10-fold diluted HAV were prepared to inoculate cells in a 96-well plate. Monolayers were then incubated for seven, 10 and 14 days and the correlation between the ELISA in situ and RT-PCR results was evaluated. At 10 days post-incubation, the highest viral load was observed in all stocks of HAV via RT-PCR (10(5) copies/mL) (p = 0.0002), while ELISA revealed the highest quantity of particles after 14 days (optical density = 0.24, p < 0.001). At seven days post-infection, there was a significant statistical correlation between the results of the two methods, indicating equivalents titres of particles and HAV genome during this period of infection. The results reported here indicate that the duration of growth of HAV in cell culture must be taken into account to correlate genome quantification with infectivity.


Asunto(s)
Animales , Virus Defectuosos/fisiología , Ensayo de Inmunoadsorción Enzimática/métodos , Virus de la Hepatitis A/fisiología , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Replicación Viral/fisiología , Línea Celular , Macaca mulatta , Factores de Tiempo , Carga Viral , Ensayo de Placa Viral
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