Author | Santos, Alcione de Oliveira dos | |
Author | Souza, Luan Felipo Botelho | |
Author | Borzacov, Lourdes Maria | |
Author | Salcedo, Juan Miguel Villalobos | |
Author | Vieira, Deusilene Souza | |
Access date | 2016-04-13T14:19:19Z | |
Available date | 2016-04-13T14:19:19Z | |
Document date | 2014 | |
Citation | SANTOS, Alcione de Oliveira dos et al. Development of cost-effective real-time PCR test: to detect a wide range of HBV DNA concentrations in the western amazon region of Brazil. Virology Journal, v. 11, p. 1-6, 2014. | pt_BR |
URI | https://www.arca.fiocruz.br/handle/icict/13747 | |
Language | eng | pt_BR |
Publisher | Virology Journal | pt_BR |
Rights | open access | pt_BR |
Title | Development of cost-effective real-time PCR test: to detect a wide range of HBV DNA concentrations in the western amazon region of Brazil | pt_BR |
Type | Article | pt_BR |
DOI | 10.1186/1743-422X-11-16 | |
Abstract | Background: Currently there is a significant risk of infection with hepatitis B virus (HBV) during blood transfusion in high epidemic area. This is due to the pre-seroconversion window period, immunovariant viral strains and the presence of occult HBV infection (OBI). The aim of this study was to develop an in-house real-time PCR-based
method, which was both ultra-sensitive and efficient offering an alternative method for nucleic acid testing (NAT).
Methods: A precore fragment with 109 bp was cloned and serial diluted to standard curve construction. The calibration of the HBV - DNA values was performed against OptiQuant® HBV-DNA Quantification Panel, Acrometrix Europe B.V.). Results: From our in-house plasmid we prepared serial dilutions ranging from 2 × 103 – 2 × 109 copies/ml. The threshold was adjusted automatically during analysis and the data collected were analyzed by linear regression (r2 = 0.99). The limit of detection for the assay with pHBVRO standards was 2000/ml in a total reaction volume of 30 μl. We found a strong correlation between the two methods (r2 = 0.9965 and p < 0.0001). The regression line give us the following equation: Log 10 (IU/mL) = 0.9038Log 10 (copies/mL) − 1.0643, suggesting that 1 IU/mL = 15
copies/mL.
Conclusions: Therefore, we can affirm that the qHBVRO PCR can detect HBV DNA in individuals with hepatitis B at any
stage of the disease showing high capacity for NAT screening in hepatitis b donors. This results of sensitivity could
provide an advance for automation in blood banks and increasing safety of patients who receive blood transfusions. | pt_BR |
Affilliation | Fundação Oswaldo Cruz. Porto Velho, RO, Brazil / Centro de Pesquisa em Medicina Tropical de Rondônia. Porto Velho, RO, Brazil / Universidade Federal de Rondônia. Porto Velho, RO, Brazil. | pt_BR |
Affilliation | Fundação Oswaldo Cruz. Porto Velho, RO, Brazil / Centro de Pesquisa em Medicina Tropical de Rondônia. Porto Velho, RO, Brazil / Universidade Federal de Rondônia. Porto Velho, RO, Brazil. | pt_BR |
Affilliation | Centro de Pesquisa em Medicina Tropical de Rondônia. Porto Velho, RO, Brazil. | pt_BR |
Affilliation | Fundação Oswaldo Cruz. Porto Velho, RO, Brazil / Centro de Pesquisa em Medicina Tropical de Rondônia. Porto Velho, RO, Brazil / Universidade Federal de Rondônia. Porto Velho, RO, Brazil. | pt_BR |
Affilliation | Fundação Oswaldo Cruz. Porto Velho, RO, Brazil / Centro de Pesquisa em Medicina Tropical de Rondônia. Porto Velho, RO, Brazil / Universidade Federal de Rondônia. Porto Velho, RO, Brazil. | pt_BR |
Subject | PCR | en |
Subject | DNA | en |
Subject | HBV | en |
Subject | Western Amazon Region | en |