Ultra-deep sequencing reveals high prevalence and broad structural diversity of hepatitis B surface antigen mutations in a global population

Mikael Gencay, Kirsten Hübner, Peter Gohl, Anja Seffner, Michael Weizenegger, Dionysios Neofytos, Richard Batrla, Andreas Woeste, Hyonsuk Kim, Gaston Westergaard, Christine Reinsch, Eva Brill, Pham Thi Thu Thuy, Bui Huu Hoang, Mark Sonderup, C. Wendy Spearman, Stephan Pabinger, Jérémie Gautier, Giuseppina Brancaccio, Massimo Fasano & 4 others Teresa Santantonio, Giovanni B. Gaeta, Markus Nauck, Wolfgang E. Kaminski

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

The diversity of the hepatitis B surface antigen (HBsAg) has a significant impact on the performance of diagnostic screening tests and the clinical outcome of hepatitis B infection. Neutralizing or diagnostic antibodies against the HBsAg are directed towards its highly conserved major hydrophilic region (MHR), in particular towards its "a" determinant subdomain. Here, we explored, on a global scale, the genetic diversity of the HBsAg MHR in a large, multi-ethnic cohort of randomly selected subjects with HBV infection from four continents. A total of 1553 HBsAg positive blood samples of subjects originating from 20 different countries across Africa, America, Asia and central Europe were characterized for amino acid variation in the MHR. Using highly sensitive ultra-deep sequencing, we found 72.8% of the successfully sequenced subjects (n = 1391) demonstrated amino acid sequence variation in the HBsAg MHR. This indicates that the global variation frequency in the HBsAg MHR is threefold higher than previously reported. The majority of the amino acid mutations were found in the HBV genotypes B (28.9%) and C (25.4%). Collectively, we identified 345 distinct amino acid mutations in the MHR. Among these, we report 62 previously unknown mutations, which extends the worldwide pool of currently known HBsAg MHR mutations by 22%. Importantly, topological analysis identified the "a" determinant upstream flanking region as the structurally most diverse subdomain of the HBsAg MHR. The highest prevalence of "a" determinant region mutations was observed in subjects from Asia, followed by the African, American and European cohorts, respectively. Finally, we found that more than half (59.3%) of all HBV subjects investigated carried multiple MHR mutations. Together, this worldwide ultra-deep sequencing based genotyping study reveals that the global prevalence and structural complexity of variation in the hepatitis B surface antigen have, to date, been significantly underappreciated.

Original languageEnglish
Article numbere0172101
JournalPloS one
Volume12
Issue number5
DOIs
Publication statusPublished - 2017 May 1

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High-Throughput Nucleotide Sequencing
Hepatitis B Surface Antigens
mutation
Mutation
Population
Amino Acids
amino acids
Central Asia
high-throughput nucleotide sequencing
hepatitis B antigens
Central America
hepatitis B
African Americans
Infection
Hepatitis B
Routine Diagnostic Tests
Central European region
neutralization
infection
genotyping

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Gencay, M., Hübner, K., Gohl, P., Seffner, A., Weizenegger, M., Neofytos, D., ... Kaminski, W. E. (2017). Ultra-deep sequencing reveals high prevalence and broad structural diversity of hepatitis B surface antigen mutations in a global population. PloS one, 12(5), [e0172101]. https://doi.org/10.1371/journal.pone.0172101
Gencay, Mikael ; Hübner, Kirsten ; Gohl, Peter ; Seffner, Anja ; Weizenegger, Michael ; Neofytos, Dionysios ; Batrla, Richard ; Woeste, Andreas ; Kim, Hyonsuk ; Westergaard, Gaston ; Reinsch, Christine ; Brill, Eva ; Thuy, Pham Thi Thu ; Hoang, Bui Huu ; Sonderup, Mark ; Spearman, C. Wendy ; Pabinger, Stephan ; Gautier, Jérémie ; Brancaccio, Giuseppina ; Fasano, Massimo ; Santantonio, Teresa ; Gaeta, Giovanni B. ; Nauck, Markus ; Kaminski, Wolfgang E. / Ultra-deep sequencing reveals high prevalence and broad structural diversity of hepatitis B surface antigen mutations in a global population. In: PloS one. 2017 ; Vol. 12, No. 5.
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abstract = "The diversity of the hepatitis B surface antigen (HBsAg) has a significant impact on the performance of diagnostic screening tests and the clinical outcome of hepatitis B infection. Neutralizing or diagnostic antibodies against the HBsAg are directed towards its highly conserved major hydrophilic region (MHR), in particular towards its {"}a{"} determinant subdomain. Here, we explored, on a global scale, the genetic diversity of the HBsAg MHR in a large, multi-ethnic cohort of randomly selected subjects with HBV infection from four continents. A total of 1553 HBsAg positive blood samples of subjects originating from 20 different countries across Africa, America, Asia and central Europe were characterized for amino acid variation in the MHR. Using highly sensitive ultra-deep sequencing, we found 72.8{\%} of the successfully sequenced subjects (n = 1391) demonstrated amino acid sequence variation in the HBsAg MHR. This indicates that the global variation frequency in the HBsAg MHR is threefold higher than previously reported. The majority of the amino acid mutations were found in the HBV genotypes B (28.9{\%}) and C (25.4{\%}). Collectively, we identified 345 distinct amino acid mutations in the MHR. Among these, we report 62 previously unknown mutations, which extends the worldwide pool of currently known HBsAg MHR mutations by 22{\%}. Importantly, topological analysis identified the {"}a{"} determinant upstream flanking region as the structurally most diverse subdomain of the HBsAg MHR. The highest prevalence of {"}a{"} determinant region mutations was observed in subjects from Asia, followed by the African, American and European cohorts, respectively. Finally, we found that more than half (59.3{\%}) of all HBV subjects investigated carried multiple MHR mutations. Together, this worldwide ultra-deep sequencing based genotyping study reveals that the global prevalence and structural complexity of variation in the hepatitis B surface antigen have, to date, been significantly underappreciated.",
author = "Mikael Gencay and Kirsten H{\"u}bner and Peter Gohl and Anja Seffner and Michael Weizenegger and Dionysios Neofytos and Richard Batrla and Andreas Woeste and Hyonsuk Kim and Gaston Westergaard and Christine Reinsch and Eva Brill and Thuy, {Pham Thi Thu} and Hoang, {Bui Huu} and Mark Sonderup and Spearman, {C. Wendy} and Stephan Pabinger and J{\'e}r{\'e}mie Gautier and Giuseppina Brancaccio and Massimo Fasano and Teresa Santantonio and Gaeta, {Giovanni B.} and Markus Nauck and Kaminski, {Wolfgang E.}",
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Gencay, M, Hübner, K, Gohl, P, Seffner, A, Weizenegger, M, Neofytos, D, Batrla, R, Woeste, A, Kim, H, Westergaard, G, Reinsch, C, Brill, E, Thuy, PTT, Hoang, BH, Sonderup, M, Spearman, CW, Pabinger, S, Gautier, J, Brancaccio, G, Fasano, M, Santantonio, T, Gaeta, GB, Nauck, M & Kaminski, WE 2017, 'Ultra-deep sequencing reveals high prevalence and broad structural diversity of hepatitis B surface antigen mutations in a global population', PloS one, vol. 12, no. 5, e0172101. https://doi.org/10.1371/journal.pone.0172101

Ultra-deep sequencing reveals high prevalence and broad structural diversity of hepatitis B surface antigen mutations in a global population. / Gencay, Mikael; Hübner, Kirsten; Gohl, Peter; Seffner, Anja; Weizenegger, Michael; Neofytos, Dionysios; Batrla, Richard; Woeste, Andreas; Kim, Hyonsuk; Westergaard, Gaston; Reinsch, Christine; Brill, Eva; Thuy, Pham Thi Thu; Hoang, Bui Huu; Sonderup, Mark; Spearman, C. Wendy; Pabinger, Stephan; Gautier, Jérémie; Brancaccio, Giuseppina; Fasano, Massimo; Santantonio, Teresa; Gaeta, Giovanni B.; Nauck, Markus; Kaminski, Wolfgang E.

In: PloS one, Vol. 12, No. 5, e0172101, 01.05.2017.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Ultra-deep sequencing reveals high prevalence and broad structural diversity of hepatitis B surface antigen mutations in a global population

AU - Gencay, Mikael

AU - Hübner, Kirsten

AU - Gohl, Peter

AU - Seffner, Anja

AU - Weizenegger, Michael

AU - Neofytos, Dionysios

AU - Batrla, Richard

AU - Woeste, Andreas

AU - Kim, Hyonsuk

AU - Westergaard, Gaston

AU - Reinsch, Christine

AU - Brill, Eva

AU - Thuy, Pham Thi Thu

AU - Hoang, Bui Huu

AU - Sonderup, Mark

AU - Spearman, C. Wendy

AU - Pabinger, Stephan

AU - Gautier, Jérémie

AU - Brancaccio, Giuseppina

AU - Fasano, Massimo

AU - Santantonio, Teresa

AU - Gaeta, Giovanni B.

AU - Nauck, Markus

AU - Kaminski, Wolfgang E.

PY - 2017/5/1

Y1 - 2017/5/1

N2 - The diversity of the hepatitis B surface antigen (HBsAg) has a significant impact on the performance of diagnostic screening tests and the clinical outcome of hepatitis B infection. Neutralizing or diagnostic antibodies against the HBsAg are directed towards its highly conserved major hydrophilic region (MHR), in particular towards its "a" determinant subdomain. Here, we explored, on a global scale, the genetic diversity of the HBsAg MHR in a large, multi-ethnic cohort of randomly selected subjects with HBV infection from four continents. A total of 1553 HBsAg positive blood samples of subjects originating from 20 different countries across Africa, America, Asia and central Europe were characterized for amino acid variation in the MHR. Using highly sensitive ultra-deep sequencing, we found 72.8% of the successfully sequenced subjects (n = 1391) demonstrated amino acid sequence variation in the HBsAg MHR. This indicates that the global variation frequency in the HBsAg MHR is threefold higher than previously reported. The majority of the amino acid mutations were found in the HBV genotypes B (28.9%) and C (25.4%). Collectively, we identified 345 distinct amino acid mutations in the MHR. Among these, we report 62 previously unknown mutations, which extends the worldwide pool of currently known HBsAg MHR mutations by 22%. Importantly, topological analysis identified the "a" determinant upstream flanking region as the structurally most diverse subdomain of the HBsAg MHR. The highest prevalence of "a" determinant region mutations was observed in subjects from Asia, followed by the African, American and European cohorts, respectively. Finally, we found that more than half (59.3%) of all HBV subjects investigated carried multiple MHR mutations. Together, this worldwide ultra-deep sequencing based genotyping study reveals that the global prevalence and structural complexity of variation in the hepatitis B surface antigen have, to date, been significantly underappreciated.

AB - The diversity of the hepatitis B surface antigen (HBsAg) has a significant impact on the performance of diagnostic screening tests and the clinical outcome of hepatitis B infection. Neutralizing or diagnostic antibodies against the HBsAg are directed towards its highly conserved major hydrophilic region (MHR), in particular towards its "a" determinant subdomain. Here, we explored, on a global scale, the genetic diversity of the HBsAg MHR in a large, multi-ethnic cohort of randomly selected subjects with HBV infection from four continents. A total of 1553 HBsAg positive blood samples of subjects originating from 20 different countries across Africa, America, Asia and central Europe were characterized for amino acid variation in the MHR. Using highly sensitive ultra-deep sequencing, we found 72.8% of the successfully sequenced subjects (n = 1391) demonstrated amino acid sequence variation in the HBsAg MHR. This indicates that the global variation frequency in the HBsAg MHR is threefold higher than previously reported. The majority of the amino acid mutations were found in the HBV genotypes B (28.9%) and C (25.4%). Collectively, we identified 345 distinct amino acid mutations in the MHR. Among these, we report 62 previously unknown mutations, which extends the worldwide pool of currently known HBsAg MHR mutations by 22%. Importantly, topological analysis identified the "a" determinant upstream flanking region as the structurally most diverse subdomain of the HBsAg MHR. The highest prevalence of "a" determinant region mutations was observed in subjects from Asia, followed by the African, American and European cohorts, respectively. Finally, we found that more than half (59.3%) of all HBV subjects investigated carried multiple MHR mutations. Together, this worldwide ultra-deep sequencing based genotyping study reveals that the global prevalence and structural complexity of variation in the hepatitis B surface antigen have, to date, been significantly underappreciated.

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