| Author | Marques, Brunna Lemos Crespo | |
| Author | Espírito-Santo, Márcia Paschoal do | |
| Author | Marques, Vanessa Alves | |
| Author | Miguel, Juliana Custódio | |
| Author | Silva, Elisangela Ferreira da | |
| Author | Nogueira, Cristiane Alves Villela | |
| Author | Lewis-Ximenez, Lia Laura | |
| Author | Lampe, Elisabeth | |
| Author | Villar, Livia Melo | |
| Access date | 2016-12-11T13:48:12Z | |
| Available date | 2016-12-11T13:48:12Z | |
| Document date | 2016 | |
| Citation | MARQUES, Brunna Lemos Crespo; et al. Evaluation of dried blood spot samples for hepatitis C virus detection and quantification. Journal of Clinical Virology, v.82, p.139-144, 2016. | pt_BR |
| ISSN | 1386-6532 | pt_BR |
| URI | https://www.arca.fiocruz.br/handle/icict/16502 | |
| Language | eng | pt_BR |
| Publisher | Elsevier | pt_BR |
| Rights | restricted access | |
| Subject in Portuguese | Ponto de sangue seco | pt_BR |
| Subject in Portuguese | Genotipagem | pt_BR |
| Subject in Portuguese | Vírus da Hepatite C | pt_BR |
| Subject in Portuguese | Otimização | pt_BR |
| Subject in Portuguese | Métodos quantitativos | pt_BR |
| Title | Evaluation of dried blood spot samples for hepatitis C virus detection and quantification | pt_BR |
| Type | Article | |
| DOI | 10.1016/j.jcv.2016.07.009 | |
| Abstract | Abstract
BACKGROUND:
Dried blood spots (DBS) could be an excellent alternative for HCV diagnosis, since it is less invasive and can be stored and transported without refrigeration.
OBJECTIVES:
The aim of this study was to optimize quantitative and qualitative methods for HCV detection in DBS.
STUDY DESIGN:
DBS and serum samples were collected from 99 subjects (59 anti-HCV/HCV RNA positive and 40 seronegative samples). Seven extraction methods and different PCR parameters were evaluated in DBS samples in the quantitative RT-PCR (qRT-PCR) developed to amplify the 5' noncoding region of HCV. A qualitative PCR for amplification of NS5B region of HCV was also valued and the nested-PCR sequenced.
RESULTS:
The qRT-PCR showed good correlation to commercial assay for HCV viral measurement in serum. To quantify HCV RNA in DBS, it was necessary to increase reverse transcriptase and cDNA concentration. HCV RNA quantification in DBS demonstrated sensitivity of 65.9%, 100% of specificity and kappa statistic of 0.65. The median viral load of DBS samples was 5.38 log10 copies/ml (minimum value=1.76 and maximum value=10.48 log10 copies/ml). HCV RNA was detected in NS5B regions and nucleotide sequences obtained in 43 serum and 11 DBS samples. The presence of the same subtype was observed in paired serum and DBS samples.
CONCLUSIONS:
In this study, it was possible to demonstrate that, despite the low sensitivity, the optimized protocol was able to determine the viral load, as well as, the infecting HCV genotype, validating the usefulness of DBS for viral load determination and molecular epidemiology studies of HCV. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Hepatites Virais. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Hepatites Virais. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Hepatites Virais. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Hepatites Virais. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Hepatites Virais. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Universidade Federal do Rio de Janeiro. Hospital Universitário Clementino Fraga Filho. Divisão de Hepatologia. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Hepatites Virais. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Hepatites Virais. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Hepatites Virais. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Subject | Dried blood spot (DBS) | pt_BR |
| Subject | Genotyping | pt_BR |
| Subject | Hepatitis C virus | pt_BR |
| Subject | Optimization | pt_BR |
| Subject | Quantitative methods | pt_BR |
| e-ISSN | 1873-5967 | |
| Embargo date | 2030-01-01 | |
