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Dried blood spot sampling for hepatitis B virus quantification, sequencing and mutation detection

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,...

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Autores principales: Bezerra, Cristianne Sousa, Portilho, Moyra Machado, Barbosa, Jakeline Ribeiro, de Azevedo, Carolina Pimentel, Mendonça, Ana Carolina da Fonseca, da Cruz, José Napoleão Monte, Frota, Cristiane Cunha, do Lago, Bárbara Vieira, Villar, Lívia Melo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8803841/
https://www.ncbi.nlm.nih.gov/pubmed/35102169
http://dx.doi.org/10.1038/s41598-022-05264-1
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author Bezerra, Cristianne Sousa
Portilho, Moyra Machado
Barbosa, Jakeline Ribeiro
de Azevedo, Carolina Pimentel
Mendonça, Ana Carolina da Fonseca
da Cruz, José Napoleão Monte
Frota, Cristiane Cunha
do Lago, Bárbara Vieira
Villar, Lívia Melo
author_facet Bezerra, Cristianne Sousa
Portilho, Moyra Machado
Barbosa, Jakeline Ribeiro
de Azevedo, Carolina Pimentel
Mendonça, Ana Carolina da Fonseca
da Cruz, José Napoleão Monte
Frota, Cristiane Cunha
do Lago, Bárbara Vieira
Villar, Lívia Melo
author_sort Bezerra, Cristianne Sousa
collection PubMed
description 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.
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spelling pubmed-88038412022-02-01 Dried blood spot sampling for hepatitis B virus quantification, sequencing and mutation detection Bezerra, Cristianne Sousa Portilho, Moyra Machado Barbosa, Jakeline Ribeiro de Azevedo, Carolina Pimentel Mendonça, Ana Carolina da Fonseca da Cruz, José Napoleão Monte Frota, Cristiane Cunha do Lago, Bárbara Vieira Villar, Lívia Melo Sci Rep Article 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. Nature Publishing Group UK 2022-01-31 /pmc/articles/PMC8803841/ /pubmed/35102169 http://dx.doi.org/10.1038/s41598-022-05264-1 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Bezerra, Cristianne Sousa
Portilho, Moyra Machado
Barbosa, Jakeline Ribeiro
de Azevedo, Carolina Pimentel
Mendonça, Ana Carolina da Fonseca
da Cruz, José Napoleão Monte
Frota, Cristiane Cunha
do Lago, Bárbara Vieira
Villar, Lívia Melo
Dried blood spot sampling for hepatitis B virus quantification, sequencing and mutation detection
title Dried blood spot sampling for hepatitis B virus quantification, sequencing and mutation detection
title_full Dried blood spot sampling for hepatitis B virus quantification, sequencing and mutation detection
title_fullStr Dried blood spot sampling for hepatitis B virus quantification, sequencing and mutation detection
title_full_unstemmed Dried blood spot sampling for hepatitis B virus quantification, sequencing and mutation detection
title_short Dried blood spot sampling for hepatitis B virus quantification, sequencing and mutation detection
title_sort dried blood spot sampling for hepatitis b virus quantification, sequencing and mutation detection
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8803841/
https://www.ncbi.nlm.nih.gov/pubmed/35102169
http://dx.doi.org/10.1038/s41598-022-05264-1
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