Hepatitis B virus genotypes and mutations in the basal
                    core promoter and pre-core/core in chronically infected patients in
                    southern Brazil: a cross-sectional study of HBV genotypes and mutations in
                    chronic carriers

Mello, Flávia Miryan Martins Almeida de; Kuniyoshi, Aline Satie Oba; Lopes, André Fanhani; Gomes-Gouvêa, Michele Soares; Bertolini, Dennis Armando

doi:10.1590/0037-8682-0158-2014

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Major Articles • Rev. Soc. Bras. Med. Trop. 47 (6) • Nov-Dec 2014 • https://doi.org/10.1590/0037-8682-0158-2014 copy

Hepatitis B virus genotypes and mutations in the basal core promoter and pre-core/core in chronically infected patients in southern Brazil: a cross-sectional study of HBV genotypes and mutations in chronic carriers

Authorship SCIMAGO INSTITUTIONS RANKINGS

Introduction

In Brazil, little data exist regarding the distribution of genotypes in relation to basal core promoter (BCP) and precore/core mutations among chronic hepatitis B virus (HBV) carriers from different regions of the country. The aim of this study was to identify HBV genotypes and the frequency of mutations at the BCP and precore/core region among the prevalent genotypes in chronic carriers from southern Brazil.

Methods

Nested-polymerase chain reaction (nested-PCR) products amplified from the S-polymerase gene, BCP and precore/core region from 54 samples were sequenced and analyzed.

Results

Phylogenetic analysis of the S-polymerase gene sequences showed that 66.7% (36/54) of the patients were infected with genotype D (D1, D2, D3), 25.9% (14/54) with genotype A (A1, A2), 5.6% (3/54) with subgenotype C2, and 2% (1/54) with genotype E. A comparison of virological characteristics showed significant differences between genotypes A, C and D. The comparison between HBeAg status and the G1896A stop codon mutation in patients with genotype D revealed a relationship between HBV G1896A precore mutants and genotype D and hepatitis B e antigen (HBeAg) seroconversion. Genotype D had a higher prevalence of the G1896A mutation and the presence of a thymine at position 1858. Genotype A was associated with a higher prevalence of the G1862T mutation and the presence of a cytosine at position 1858.

Conclusions

HBV genotype D (D3) is predominant in HBV chronic carriers from southern Brazil. The presence of mutations in the BCP and precore/core region was correlated with the HBV genotype and HBeAg negative status.

HBV; Genotypes; Subgenotypes; Basal core promoter mutations; Precore/core mutations

Characteristic	A	C	D	p
Characteristic	(n=14)	(n=3)	(n=36)	p
Age (years)
mean ± SD	42.7±10.6	48±8.5	42±12.95	0.712#
range	(31-70)	(39-56)	(20-67)
median	42.0	49.0	41.5
Gender (male/female)	9/5	2/1	18/18	0.646^§ § Fisher's exact test; Note: The data from the only patient infected with genotype E are not present in the table. HBV: hepatitis B virus; SD: standard deviation; HBeAg: hepatitis B e antigen; ALT: alanine aminotransferase; AST: aspartate aminotransferase; ANOVA: analysis of variance.
HBeAg-positive patients, n (%)	3 (21.4%)	1 (33.3%)	4 (11.1%)	0.304^§ § Fisher's exact test; Note: The data from the only patient infected with genotype E are not present in the table. HBV: hepatitis B virus; SD: standard deviation; HBeAg: hepatitis B e antigen; ALT: alanine aminotransferase; AST: aspartate aminotransferase; ANOVA: analysis of variance.
Indicators of liver function
elevated ALT, n (%)^* * When higher than 2 times the upper limit of normal;	2 (14.0%)	1 (33.3%)	7 (19.4%)	0.702^§ § Fisher's exact test; Note: The data from the only patient infected with genotype E are not present in the table. HBV: hepatitis B virus; SD: standard deviation; HBeAg: hepatitis B e antigen; ALT: alanine aminotransferase; AST: aspartate aminotransferase; ANOVA: analysis of variance.
elevated AST, n (%)^* * When higher than 2 times the upper limit of normal;	3 (21.4%)	1 (33.3%)	8 (22.2%)	0.871^§ § Fisher's exact test; Note: The data from the only patient infected with genotype E are not present in the table. HBV: hepatitis B virus; SD: standard deviation; HBeAg: hepatitis B e antigen; ALT: alanine aminotransferase; AST: aspartate aminotransferase; ANOVA: analysis of variance.
Patients with viral mutations, n (%)
T1753V	5 (35.7%)	1 (33.3%)	12 (33.3%)	1.000^§ § Fisher's exact test; Note: The data from the only patient infected with genotype E are not present in the table. HBV: hepatitis B virus; SD: standard deviation; HBeAg: hepatitis B e antigen; ALT: alanine aminotransferase; AST: aspartate aminotransferase; ANOVA: analysis of variance.
A1762T G1764A	9 (64.3%)	2 (66.7%)	11 (30.5%)	0.050^§ § Fisher's exact test; Note: The data from the only patient infected with genotype E are not present in the table. HBV: hepatitis B virus; SD: standard deviation; HBeAg: hepatitis B e antigen; ALT: alanine aminotransferase; AST: aspartate aminotransferase; ANOVA: analysis of variance.
G1896A	0 (0.0%)	1 (33.3%)	28 (77.7%)	<0.001^§ § Fisher's exact test; Note: The data from the only patient infected with genotype E are not present in the table. HBV: hepatitis B virus; SD: standard deviation; HBeAg: hepatitis B e antigen; ALT: alanine aminotransferase; AST: aspartate aminotransferase; ANOVA: analysis of variance.
C1858T	3 (21.4%)	3 (100.0%)	36 (100.0%)	<0.001^§ § Fisher's exact test; Note: The data from the only patient infected with genotype E are not present in the table. HBV: hepatitis B virus; SD: standard deviation; HBeAg: hepatitis B e antigen; ALT: alanine aminotransferase; AST: aspartate aminotransferase; ANOVA: analysis of variance.
G1899A	1 (7.1%)	1 (33.3%)	12 (33.3%)	0.122^§ § Fisher's exact test; Note: The data from the only patient infected with genotype E are not present in the table. HBV: hepatitis B virus; SD: standard deviation; HBeAg: hepatitis B e antigen; ALT: alanine aminotransferase; AST: aspartate aminotransferase; ANOVA: analysis of variance.
G1862T	9 (64.0%)	0 (0.0%)	2 (5.5%)	<0.001^§ § Fisher's exact test; Note: The data from the only patient infected with genotype E are not present in the table. HBV: hepatitis B virus; SD: standard deviation; HBeAg: hepatitis B e antigen; ALT: alanine aminotransferase; AST: aspartate aminotransferase; ANOVA: analysis of variance.

Sociedade Brasileira de Medicina Tropical - SBMT Sociedade Brasileira de Medicina Tropical - SBMT, Núcleo de Medicina Tropical UnB, Sala 43C 70904-970, E-mails: rsbmt@uftm.edu.br | artes.rsbmt@gmail.com | sbmt@sbmt.org.br , WhatsApp: SBMT (61) 9.9192-6496, WhatsApp: RSBMT (34) 9.9996-5807 - Brasília - DF - Brazil
E-mail: rsbmt@uftm.edu.br

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[1] Address to: Dr. Dennis Armando Bertolini. Laboratório de Virologia Clínica/Depto. de Análises Clínicas e Biomedicina/UEM. Av. Colombo 5790, 87020-900 Maringá, PR, Brasil. Phone: 55 44 3011-5394; Fax: 55 44 3011-4860. e-mail: dabertolini@uem.br