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Applicability of oral fluid samples for tracking hepatitis B virus mutations, genotyping, and phylogenetic analysis

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Abstract

Little is known about the usefulness of saliva samples for hepatitis B virus (HBV) genotyping and mutation analysis. The aim of this study was to evaluate the usefulness of oral fluid samples to determine HBV genotype distribution, S/polymerase mutations, and HBV subpopulation diversity among chronically HBV-infected individuals. Serum and oral fluid samples were obtained from 18 individuals for PCR and nucleotide sequencing of the HBV surface antigen gene. Biochemical analysis of liver enzymes (ALT, AST, GGT) and HBV, HCV, and HIV serological tests were also performed. All serum samples were HBsAg (+), anti-HBc (+), and anti-HBs (−); 55.6% were HBeAg (+)/anti-HBe (−), and 11.1% were anti-HIV (+). The mean HBV DNA viral load was 6.1 ± 2.3 log IU/mL. The HBV genotype distribution was as follows: A, 72.2%; D, 11.1%; E, 5.6%; F, 11.1%. A concordance of 100% in genotype classification and 99.8% in sequence similarity between paired oral fluid and serum samples was observed. HBsAg mutations were detected in all samples, but no resistance mutations were found in the polymerase gene. This study demonstrates that oral fluid samples can be used reliably for tracking HBV mutations, genotyping, and phylogenetic analysis. This could be important for molecular epidemiology studies with hard-to-reach populations.

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Availability of data and material

The datasets generated and/or analysed during the current study are not publicly available, in order to maintain the privacy and confidentiality of the subjects, but are available from the corresponding author upon reasonable request.

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Acknowledgements

The authors wish to acknowledge the support of staff members of the public health centers Viral Hepatitis Ambulatory/Laboratory (IOC/Fiocruz, Rio de Janeiro), Clementino Fraga Filho Hospital (Federal University of Rio de Janeiro, Rio de Janeiro), and Health Unit (Center of Guidance and Advisor, COA, Curitiba), who worked in sample collection and patient assistance.

Funding

This study was supported by Oswaldo Cruz IOC/Fiocruz/Ministry of Health, Brazil, and the Brazilian national funding agencies Conselho Nacional de Desenvolvimento Científico e Tecnológico of Brazil (CNPq), Fundação Carlos Chagas de Amparo a Pesquisa do Estado do Rio de Janeiro (FAPERJ), and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior of Brasil (CAPES).

Author information

Authors and Affiliations

  1. Laboratory of Viral Hepatitis, Oswaldo Cruz Institute, FIOCRUZ, Helio and Peggy, Ground Floor, Room B09, Av. Brasil, 4365, Manguinhos, Rio de Janeiro, RJ, 210360-040, Brazil

    Moyra Machado Portilho, Cristianne Sousa Bezerra, Ana Carolina da Fonseca Mendonça, Vanessa Alves Marques, Lia Laura Lewis-Ximenez, Bárbara Vieira do Lago & Livia Melo Villar

  2. Gonçalo Moniz Research Institute, FIOCRUZ, Salvador, BA, Brazil

    Moyra Machado Portilho

  3. Federal Institute of Education, Science and Technology, Fortaleza, CE, Brazil

    Cristianne Sousa Bezerra

  4. Clementino Fraga Filho Hospital, University of Rio de Janeiro, UFRJ, Rio de Janeiro, RJ, Brazil

    Leticia Cancella Nabuco & Cristiane Alves Villela-Nogueira

  5. Center of Guidance and Advisor (COA), Curitiba, PR, Brazil

    Cláudia Alexandra Pontes Ivantes

  6. Immunobiological Technology Institute (Bio-Manguinhos), FIOCRUZ, Rio de Janeiro, RJ, Brazil

    Bárbara Vieira do Lago

Authors
  1. Moyra Machado Portilho
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  9. Bárbara Vieira do Lago
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  10. Livia Melo Villar
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Contributions

MMP, BVL, and LMV prepared and revised the manuscript. MMP, CSB, and ACFM did molecular tests, including sequencing. BVL and MMP performed the phylogenetics analysis. LLLX, CAVN, LCN, and CAPI contributed to recruiting and medical care of patients. VAM, CSB, MMP, and ACFM worked on data curation, performed the molecular assays, also providing the interpretation of the molecular results obtained. LMV supervised and managed the project and worked on funding acquisition. All authors revised the final version of this manuscript.

Corresponding author

Correspondence to Livia Melo Villar.

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Conflict of interest

The authors declare no conflict of interest.

Ethical approval

This study was approved by the Fiocruz Ethics Committee (number CAAE 18281313.4.0000.5248).

Consent to participate

All individuals gave formal consent to participate in this study.

Additional information

Handling Editor: Michael Carpenter.

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Institution at which the work was performed: Laboratory of Viral Hepatitis, Oswaldo Cruz Institute, FIOCRUZ, Rio de Janeiro.

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Portilho, M.M., Bezerra, C.S., Mendonça, A.C.d. et al. Applicability of oral fluid samples for tracking hepatitis B virus mutations, genotyping, and phylogenetic analysis. Arch Virol 166, 2435–2442 (2021). https://doi.org/10.1007/s00705-021-05122-x

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  • DOI: https://doi.org/10.1007/s00705-021-05122-x

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