Cargando…

Analytical validation of hepatitis B core‐related antigen (HBcrAg) using dried blood spots (DBS)

Limited access to nucleic acid testing (NAT) to quantify HBV DNA levels, an essential tool to determine anti‐HBV treatment eligibility, represents a significant barrier to scale up HBV diagnostic services in resource‐limited countries. Hepatitis B core‐related antigen (HBcrAg) has the potential to b...

Descripción completa

Detalles Bibliográficos
Autores principales: Shimakawa, Yusuke, Vernoux, Laura, Gabassi, Audrey, Mercier‐Delarue, Séverine, Vincent, Jeanne Perpétue, Simon, François, Maylin, Sarah
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8247985/
https://www.ncbi.nlm.nih.gov/pubmed/33599049
http://dx.doi.org/10.1111/jvh.13489
_version_ 1783716628658126848
author Shimakawa, Yusuke
Vernoux, Laura
Gabassi, Audrey
Mercier‐Delarue, Séverine
Vincent, Jeanne Perpétue
Simon, François
Maylin, Sarah
author_facet Shimakawa, Yusuke
Vernoux, Laura
Gabassi, Audrey
Mercier‐Delarue, Séverine
Vincent, Jeanne Perpétue
Simon, François
Maylin, Sarah
author_sort Shimakawa, Yusuke
collection PubMed
description Limited access to nucleic acid testing (NAT) to quantify HBV DNA levels, an essential tool to determine anti‐HBV treatment eligibility, represents a significant barrier to scale up HBV diagnostic services in resource‐limited countries. Hepatitis B core‐related antigen (HBcrAg) has the potential to become an affordable alternative because of its low cost (US$ <15/assay) and strong correlation with HBV DNA levels in treatment‐naïve patients. However, the current assay requires plasma or serum. To further facilitate its application to decentralized settings, we developed and evaluated a standardized procedure to quantify HBcrAg using dried blood spots as a tool to diagnose HBV‐infected people with high viraemia. We evaluated the following elution method optimized to quantify HBcrAg: suspension of a punched blood‐soaked disc (11 mm) of Whatman 903 Protein Saver Card in 450 µL of PBS 0.05% Tween 20, followed by an incubation for 4 h at room temperature and a centrifugation at 10,000 g for 10 minutes. 150 µL of DBS eluate was used to quantify HBcrAg using chemiluminescent enzyme immunoassay (LUMIPULSE(®) G600II, Fujirebio). The limit of detection of dried blood spot HBcrAg in relation with HBV DNA levels was 19,115 IU/mL across the five major HBV genotypes (A/B/C/D/E). A strong linear correlation was confirmed between dried blood spot HBcrAg and HBV DNA levels (r = 0.94, p < 0.0001) in samples with high viral loads (range: 3.7–7.0 log IU/mL). The coefficient of variation ranged between 4.0–11.2% for repeatability and 3.9–12.2% for reproducibility. Analytical specificity was 100% (95% CI: 83.9–100%) in HBV‐negative samples. Using our elution method, it may be possible to identify HBV‐infected patients with high viraemia who need antiviral therapy using dried blood spot and HBcrAg. A large‐scale clinical validation is warranted in resource‐limited countries.
format Online
Article
Text
id pubmed-8247985
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher John Wiley and Sons Inc.
record_format MEDLINE/PubMed
spelling pubmed-82479852021-07-02 Analytical validation of hepatitis B core‐related antigen (HBcrAg) using dried blood spots (DBS) Shimakawa, Yusuke Vernoux, Laura Gabassi, Audrey Mercier‐Delarue, Séverine Vincent, Jeanne Perpétue Simon, François Maylin, Sarah J Viral Hepat Original Articles Limited access to nucleic acid testing (NAT) to quantify HBV DNA levels, an essential tool to determine anti‐HBV treatment eligibility, represents a significant barrier to scale up HBV diagnostic services in resource‐limited countries. Hepatitis B core‐related antigen (HBcrAg) has the potential to become an affordable alternative because of its low cost (US$ <15/assay) and strong correlation with HBV DNA levels in treatment‐naïve patients. However, the current assay requires plasma or serum. To further facilitate its application to decentralized settings, we developed and evaluated a standardized procedure to quantify HBcrAg using dried blood spots as a tool to diagnose HBV‐infected people with high viraemia. We evaluated the following elution method optimized to quantify HBcrAg: suspension of a punched blood‐soaked disc (11 mm) of Whatman 903 Protein Saver Card in 450 µL of PBS 0.05% Tween 20, followed by an incubation for 4 h at room temperature and a centrifugation at 10,000 g for 10 minutes. 150 µL of DBS eluate was used to quantify HBcrAg using chemiluminescent enzyme immunoassay (LUMIPULSE(®) G600II, Fujirebio). The limit of detection of dried blood spot HBcrAg in relation with HBV DNA levels was 19,115 IU/mL across the five major HBV genotypes (A/B/C/D/E). A strong linear correlation was confirmed between dried blood spot HBcrAg and HBV DNA levels (r = 0.94, p < 0.0001) in samples with high viral loads (range: 3.7–7.0 log IU/mL). The coefficient of variation ranged between 4.0–11.2% for repeatability and 3.9–12.2% for reproducibility. Analytical specificity was 100% (95% CI: 83.9–100%) in HBV‐negative samples. Using our elution method, it may be possible to identify HBV‐infected patients with high viraemia who need antiviral therapy using dried blood spot and HBcrAg. A large‐scale clinical validation is warranted in resource‐limited countries. John Wiley and Sons Inc. 2021-03-01 2021-05 /pmc/articles/PMC8247985/ /pubmed/33599049 http://dx.doi.org/10.1111/jvh.13489 Text en © 2021 The Authors. Journal of Viral Hepatitis published by John Wiley & Sons Ltd. https://creativecommons.org/licenses/by/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Original Articles
Shimakawa, Yusuke
Vernoux, Laura
Gabassi, Audrey
Mercier‐Delarue, Séverine
Vincent, Jeanne Perpétue
Simon, François
Maylin, Sarah
Analytical validation of hepatitis B core‐related antigen (HBcrAg) using dried blood spots (DBS)
title Analytical validation of hepatitis B core‐related antigen (HBcrAg) using dried blood spots (DBS)
title_full Analytical validation of hepatitis B core‐related antigen (HBcrAg) using dried blood spots (DBS)
title_fullStr Analytical validation of hepatitis B core‐related antigen (HBcrAg) using dried blood spots (DBS)
title_full_unstemmed Analytical validation of hepatitis B core‐related antigen (HBcrAg) using dried blood spots (DBS)
title_short Analytical validation of hepatitis B core‐related antigen (HBcrAg) using dried blood spots (DBS)
title_sort analytical validation of hepatitis b core‐related antigen (hbcrag) using dried blood spots (dbs)
topic Original Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8247985/
https://www.ncbi.nlm.nih.gov/pubmed/33599049
http://dx.doi.org/10.1111/jvh.13489
work_keys_str_mv AT shimakawayusuke analyticalvalidationofhepatitisbcorerelatedantigenhbcragusingdriedbloodspotsdbs
AT vernouxlaura analyticalvalidationofhepatitisbcorerelatedantigenhbcragusingdriedbloodspotsdbs
AT gabassiaudrey analyticalvalidationofhepatitisbcorerelatedantigenhbcragusingdriedbloodspotsdbs
AT mercierdelarueseverine analyticalvalidationofhepatitisbcorerelatedantigenhbcragusingdriedbloodspotsdbs
AT vincentjeanneperpetue analyticalvalidationofhepatitisbcorerelatedantigenhbcragusingdriedbloodspotsdbs
AT simonfrancois analyticalvalidationofhepatitisbcorerelatedantigenhbcragusingdriedbloodspotsdbs
AT maylinsarah analyticalvalidationofhepatitisbcorerelatedantigenhbcragusingdriedbloodspotsdbs