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.

Details

Title
Dried blood spot sampling for hepatitis B virus quantification, sequencing and mutation detection
Author
Bezerra Cristianne Sousa 1 ; Portilho Moyra Machado 2 ; Barbosa, Jakeline Ribeiro 3 ; de Azevedo Carolina Pimentel 3 ; Mendonça Ana Carolina da Fonseca 3 ; da Cruz José Napoleão Monte 4 ; Frota Cristiane Cunha 5 ; do Lago Bárbara Vieira 3 ; Villar, Lívia Melo 3 

 Fundação Oswaldo Cruz, FIOCRUZ, Laboratorio de Hepatites Virais, Instituto Oswaldo Cruz, Helio and Peggy Pereira Pavillion - Ground Floor - Room B09, Rio de Janeiro, Brazil (GRID:grid.418068.3) (ISNI:0000 0001 0723 0931); Ciência E Tecnologia Do Ceará, Departamento de Educação, Instituto Federal de Educação, Fortaleza, Brazil (GRID:grid.461960.c) (ISNI:0000 0000 9352 6714) 
 FIOCRUZ, Instituto de Pesquisas Gonçalo Moniz, Salvador, Brazil (GRID:grid.418068.3) (ISNI:0000 0001 0723 0931) 
 Fundação Oswaldo Cruz, FIOCRUZ, Laboratorio de Hepatites Virais, Instituto Oswaldo Cruz, Helio and Peggy Pereira Pavillion - Ground Floor - Room B09, Rio de Janeiro, Brazil (GRID:grid.418068.3) (ISNI:0000 0001 0723 0931) 
 Laboratório Central Do Estado Do Ceará, Fortaleza, Brazil (GRID:grid.418068.3) 
 Universidade Federal Do Ceará, Departamento de Patologia E Medicina Legal, Faculdade de Medicina, Fortaleza, Brazil (GRID:grid.8395.7) (ISNI:0000 0001 2160 0329) 
Publication year
2022
Publication date
2022
Publisher
Nature Publishing Group
e-ISSN
20452322
Source type
Scholarly Journal
Language of publication
English
ProQuest document ID
2624041721
Copyright
© The Author(s) 2022. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.