Development of cost-effective real-time PCR test: to detect a wide range of HBV DNA concentrations in the western amazon region of Brazil
| Autor(a) principal: | |
|---|---|
| Data de Publicação: | 2014 |
| Outros Autores: | , , , |
| Tipo de documento: | Artigo |
| Idioma: | eng |
| Título da fonte: | Repositório Institucional da FIOCRUZ (ARCA) |
| DOI: | 10.1186/1743-422X-11-16 |
| Texto Completo: | https://www.arca.fiocruz.br/handle/icict/13747 |
Resumo: | 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. |
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Santos, Alcione de Oliveira dosSouza, Luan Felipo BotelhoBorzacov, Lourdes MariaSalcedo, Juan Miguel VillalobosVieira, Deusilene Souza2016-04-13T14:19:19Z2016-04-13T14:19:19Z2014SANTOS, 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.https://www.arca.fiocruz.br/handle/icict/1374710.1186/1743-422X-11-16engVirology JournalDevelopment of cost-effective real-time PCR test: to detect a wide range of HBV DNA concentrations in the western amazon region of Brazilinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleFundação 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.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.Centro de Pesquisa em Medicina Tropical de Rondônia. Porto Velho, RO, Brazil.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.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.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.PCRDNAHBVWestern Amazon Regioninfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZLICENSElicense.txtlicense.txttext/plain; charset=utf-82991https://www.arca.fiocruz.br/bitstream/icict/13747/1/license.txt5a560609d32a3863062d77ff32785d58MD51ORIGINALDevelopment of cost-effective.pdfDevelopment of cost-effective.pdfapplication/pdf268008https://www.arca.fiocruz.br/bitstream/icict/13747/2/Development%20of%20cost-effective.pdf97a9f9602cf50350dc69a9b2ca0b2367MD52TEXTDevelopment of cost-effective.pdf.txtDevelopment of cost-effective.pdf.txtExtracted texttext/plain28573https://www.arca.fiocruz.br/bitstream/icict/13747/3/Development%20of%20cost-effective.pdf.txt925e565223a9234a7a3f2555797222f7MD53icict/137472018-09-10 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| dc.title.pt_BR.fl_str_mv |
Development of cost-effective real-time PCR test: to detect a wide range of HBV DNA concentrations in the western amazon region of Brazil |
| 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 |
| spellingShingle |
Development of cost-effective real-time PCR test: to detect a wide range of HBV DNA concentrations in the western amazon region of Brazil Santos, Alcione de Oliveira dos PCR DNA HBV Western Amazon Region |
| title_short |
Development of cost-effective real-time PCR test: to detect a wide range of HBV DNA concentrations in the western amazon region of Brazil |
| title_full |
Development of cost-effective real-time PCR test: to detect a wide range of HBV DNA concentrations in the western amazon region of Brazil |
| title_fullStr |
Development of cost-effective real-time PCR test: to detect a wide range of HBV DNA concentrations in the western amazon region of Brazil |
| title_full_unstemmed |
Development of cost-effective real-time PCR test: to detect a wide range of HBV DNA concentrations in the western amazon region of Brazil |
| title_sort |
Development of cost-effective real-time PCR test: to detect a wide range of HBV DNA concentrations in the western amazon region of Brazil |
| author |
Santos, Alcione de Oliveira dos |
| author_facet |
Santos, Alcione de Oliveira dos Souza, Luan Felipo Botelho Borzacov, Lourdes Maria Salcedo, Juan Miguel Villalobos Vieira, Deusilene Souza |
| author_role |
author |
| author2 |
Souza, Luan Felipo Botelho Borzacov, Lourdes Maria Salcedo, Juan Miguel Villalobos Vieira, Deusilene Souza |
| author2_role |
author author author author |
| dc.contributor.author.fl_str_mv |
Santos, Alcione de Oliveira dos Souza, Luan Felipo Botelho Borzacov, Lourdes Maria Salcedo, Juan Miguel Villalobos Vieira, Deusilene Souza |
| dc.subject.en.en.fl_str_mv |
PCR DNA HBV Western Amazon Region |
| topic |
PCR DNA HBV Western Amazon Region |
| description |
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. |
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2014 |
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2014 |
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2016-04-13T14:19:19Z |
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2016-04-13T14:19:19Z |
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info:eu-repo/semantics/publishedVersion |
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info:eu-repo/semantics/article |
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article |
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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. |
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https://www.arca.fiocruz.br/handle/icict/13747 |
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10.1186/1743-422X-11-16 |
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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. 10.1186/1743-422X-11-16 |
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Virology Journal |
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Virology Journal |
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