| Author | Bezerra, Cristianne Sousa | |
| Author | Portilho, Moyra Machado | |
| Author | Barbosa, Jakeline Ribeiro | |
| Author | Azevedo, Carolina Pimentel de | |
| Author | Mendonça, Ana Carolina da Fonseca | |
| Author | Cruz, José Napoleão Monte da | |
| Author | Frota, Cristiane Cunha | |
| Author | Lago, Bárbara Vieira do | |
| Author | Villar, Lívia Melo | |
| Access date | 2022-08-16T13:23:22Z | |
| Available date | 2022-08-16T13:23:22Z | |
| Document date | 2022 | |
| Citation | BEZERRA, Cristianne Sousa et al. Dried blood spot sampling for hepatitis B virus quantification, sequencing and mutation detection. Scientifc Reports, v. 12, 1651, p. 1 - 11, 2022. | pt_BR |
| ISSN | 2045-2322 | pt_BR |
| URI | https://www.arca.fiocruz.br/handle/icict/54656 | |
| Language | eng | pt_BR |
| Publisher | Nature Portfolio | pt_BR |
| Rights | open access | |
| Subject in Portuguese | Amostragem de sangue seco | pt_BR |
| Subject in Portuguese | Quantificação | pt_BR |
| Subject in Portuguese | Vírus da hepatite B | pt_BR |
| Subject in Portuguese | Sequenciamento e detecção | pt_BR |
| Subject in Portuguese | Mutações | pt_BR |
| Title | Dried blood spot sampling for hepatitis B virus quantification, sequencing and mutation detection | pt_BR |
| Type | Article | |
| DOI | 10.1038/s41598-022-05264-1 | |
| Abstract | Hepatitis B virus (HBV) diagnosis is performed on serum samples, but the access to this diagnosis is
difficult in low-income regions. The use of dried blood spot (DBS) samples does not require special
structure for collection, storage or transport. This study evaluates the use of DBS for detection,
quantification and sequencing of HBV DNA using in-house techniques. Two study groups were
included: 92 HBsAg + individuals and 49 negative controls. Serum and DBS samples were submitted
to quantitative and qualitative in-house PCR for S/pol genes, sequencing and phylogenetic analyses.
Total of 84 serum samples were successfully amplified. Of them, 63 paired DBS were also positive
in qualitative PCR. Qualitative PCR in DBS presented a sensitivity of 75% and specificity of 100%
(Kappa = 0.689). Quantitative PCR in DBS presented a detection limit of 852.5 copies/mL (250 IU/mL),
sensitivity of 77.63% and specificity of 100% (Kappa = 0.731). A total of 63 serum samples and 36 DBS
samples were submitted to sequencing, revealing the circulation of genotypes A (65.08%), D (4.8%),
E (3.2%) and F (27%) with 100% of correspondence between serum and DBS. All sequenced samples
displayed polymorphisms in HBsAg gene. An HIV-coinfected patient presented the rtM204V/I-rtL180M
double resistance mutation in serum and DBS. In conclusion, DBS is an alternative to detect, quantify
and characterize HBV DNA, being a possibility of increasing diagnosis in low-income settings, closing
gaps in HBV control. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Hepatitis Virais. Rio de Janeiro, RJ, Brasil / Instituto Federal de Educação, Ciência e Tecnologia do Ceará. Departamento de Educação. Fortaleza, CE, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Hepatitis Virais. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Hepatitis Virais. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Hepatitis Virais. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Laboratório Central do Estado do Ceará. Fortaleza, CE, Brasil. | pt_BR |
| Affilliation | Universidade Federal do Ceará. Faculade de Medicina. Departamento de Patologia e Medicina Legal. Fortaleza, CE, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Hepatitis Virais. Rio de Janeiro, RJ, Brasil | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Hepatitis Virais. Rio de Janeiro, RJ, Brasil | pt_BR |
| Subject | Dried blood spot sampling | pt_BR |
| Subject | Hepatitis B virus | pt_BR |
| Subject | Quantification | pt_BR |
| Subject | Sequencing | pt_BR |
| Subject | Mutation detection | pt_BR |
