In-house quantitative real-time PCR for the diagnosis of hepatitis B virus and hepatitis C virus infections
Zauli, Danielle Alves Gomes; Menezes, Carla Lisandre Paula de; Oliveira, Cristiane Lommez de; Mateo, Elvis Cristian Cueva; Ferreira, Alessandro Clayton de Souza.
Braz. J. Microbiol.;
47(4): 987-992, Out-Dez. 2016. tab, ilus, graf
Artigo
em Inglês
| VETINDEX
| ID: vti-23303
Resumo
The quantification of viral nucleic acids in serum by real-time PCR plays an important role in diagnosing hepatitis B virus and hepatitis C virus infection. In this study, we developed an assay using specific primers and probes to quantify hepatitis B virus DNA or hepatitis C virus RNA in serum from infected patients. For standardization and validation of the assay, an international panel of hepatitis B virus/hepatitis C virus and standard plasmids was used. A correlation coefficient of 0.983 and 0.963 for hepatitis B virus and hepatitis C virus, respectively, was obtained based on cycle threshold values and concentrations of DNA or RNA. The standard curve showed a linear relationship from 19 IU/mL to 1.9 × 109 IU/mL of serum, with a coefficient of determination (r2) of 0.99. In sera from patients infected with hepatitis B virus or hepatitis C virus viral loads (19 IU/mL and 1.9 × 109 IU/mL), we quantified viral loads with a detection limit of 1.9 × 102 IU/mL. The real-time quantitative PCR assay developed in this study provides an ideal system for routine diagnosis and confirmation of indeterminate serological results, especially in immunosuppressed patients.(AU)
Biblioteca responsável:
BR68.1
Localização: BR68.1
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