Detection of replication-defective hepatitis A virus based on the correlation between real-time polymerase chain reaction and ELISA in situ results

dc.creatorCosta, Alyne Moraes
dc.creatorAmado, Luciane Almeida
dc.creatorPaula, Vanessa Salete de
dc.date2015-09-21T17:25:27Z
dc.date2015-09-21T17:25:27Z
dc.date2013
dc.date.accessioned2023-09-26T20:34:52Z
dc.date.available2023-09-26T20:34:52Z
dc.identifierCOSTA, Alyne Moraes; AMADO, Luciane Almeida; PAULA, Vanessa Salete de. Detection of replication-defective hepatitis A virus based on the correlation between real-time polymerase chain reaction and ELISA in situ results. Mem Inst Oswaldo Cruz, v.108, n.1, p.36-40, Feb. 2013.
dc.identifier1678-8060
dc.identifierhttps://www.arca.fiocruz.br/handle/icict/11717
dc.identifier.urihttps://repositorioslatinoamericanos.uchile.cl/handle/2250/8859962
dc.descriptionELISA 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 (105 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.
dc.formatapplication/pdf
dc.languageeng
dc.publisherFundação Oswaldo Cruz
dc.rightsopen access
dc.subjectHepatitis A virus
dc.subjectReplication-defective
dc.subjectRT-PCR
dc.subjectELISA in situ
dc.subjectVírus da Hepatite A
dc.subjectEnsaio de Imunoadsorção Enzimática
dc.titleDetection of replication-defective hepatitis A virus based on the correlation between real-time polymerase chain reaction and ELISA in situ results
dc.typeArticle


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